CN102123712B - Methods of cancer treatment with IGF1R inhibitors - Google Patents

Abstract

This invention relates to compositions and methods useful for treating various cancers. Therapeutic combinations and methods of use thereof are also covered in the present application.

Description

Use the method for IGF1R inhibitor for treating cancer

The application requires the U.S. Provisional Patent Application number 60/874 of December in 2006 application on the 13rd, 589, the U.S. Provisional Patent Application of December in 2006 application on the 20th number 60/870,937, the U.S. Provisional Patent Application of on June 25th, 2007 application number 60/946, the rights and interests of the U.S. Provisional Patent Application of on October 11st, 011 and 2007 application number 60/979,274; Described each temporary patent application is all attached to herein by reference.

Invention field

The present invention relates to be used for the treatment of or compositions and the method for prophylaxis of cancer.

Background of invention

Insulin like growth factor, also claim somatomedin, comprise insulin like growth factor-1 (IGF-I) and insulin like growth factor-1 I (IGF-II) ((1978) Febs.Lett.89:283 such as (1983) Endocrinol.112:2215 such as Klapper and Rinderknecht).These somatomedin are by the common receptors bind with being called IGF-1R (IGF1R or IGFR1), to comprising tumor cell (Macaulay, (1992) Br.J.Cancer 65:311) at interior various cell type performance mitogenic activities (Sepp-Lorenzino, (1998) Breast CancerResearch and Treatment 47:235).The interaction of IGF and IGF1R is activated receptor (Butler etc. (1998) ComparativeBiochemistry and Physiology 121:19) by the autophosphorylation of initiation receptor tyrosine residue.Once IGF1R is activated, even if with making in born of the same parents target phosphorylation with active cell signal transduction pathway.The activation of this receptor is very important to Tumor Cell Growth Stimulated and survival.Therefore, suppress the potential method that has much value that IGF1R activity is representing treatment or the growth of prevention human carcinomas and other proliferative disease.

Therefore, the therapy of inhibition IGF1R can be used for treatment or prevents some cancer.Anti-IGF1R antibody is to be used for the treatment of or the effective therapy of prophylaxis of cancer.Known in the art have several anti-IGF1R antibody (referring to for example WO 03/100008, WO 2002/53596, WO 04/71529, WO03/106621, US2003/235582, WO 04/83248, WO 03/59951, WO 04/87756 or WO 2005/16970).Other micromolecule IGF 1R inhibitor is also known in the art.

Although exist known in the art can be used for treat or prevent the IGF1R inhibitor of some cancer, this area still needs to be used for the treatment of or to prevent therapeutic combination and the method for other cancers such as incidence cancer, squamous cell carcinoma, solitary plasmacytoma, multiple myeloma and renal cell carcinoma.

Summary of the invention

The present invention is by effective treatment being provided or preventing IGF1R inhibitor and the combination product thereof of following disease, and part has met this needs: incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma (EwingSarcoma), chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin (Hodgekin ' sdisease), non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, marginal zone lymphoma (marginal zone lymphoma), Burkitt lymphoma, mycosis fungoides, seary's syndrome (seary syndrome), cutaneous T cell lymphoma, lymphoma peripheral T cell, chronic myeloproliferative diseases, myelofibrosis, myeloid metaplasia, systemic mastocytosis) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor or hepatocarcinoma.

The invention provides the method that is selected from following medical conditions that is used for the treatment of or prevents curee, the method comprises and gives one or more IGF1R inhibitor or its pharmaceutical composition that described curee treats effective dose: incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma, chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, Burkitt lymphoma, mycosis fungoides, seary's syndrome, cutaneous T cell lymphoma, chronic myeloproliferative diseases) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor and hepatocarcinoma.In one embodiment of the invention, IGF1R inhibitor is selected from

with for example, separation antibody with IGF1R (people IGF1R) or its Fab specific binding.In one embodiment of the invention, antibody comprises: the variable region of light chain of aminoacid 20-128 and the variable region of heavy chain of the aminoacid 20-137 that comprises SEQ ID NO:10 or 12 that (a) comprise SEQ ID NO:2; (b) variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that comprises SEQ ID NO:4 and the aminoacid 20-137 that comprises SEQID NO:10 or 12; (c) variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that comprises SEQ ID NO:6 and the aminoacid 20-137 that comprises SEQ ID NO:10 or 12; (d) variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that comprises SEQ ID NO:8 and the aminoacid 20-137 that comprises SEQ ID NO:10 or 12; Or any other IGF1R inhibitor shown in this article, for example any other IGF1R inhibitor shown in " IGF1R inhibitor " part below.In one embodiment of the invention, the IGF1R inhibitor anticancer chemotherapeutic agent other with one or more or its pharmaceutical composition are combined and are given.In one embodiment of the invention, other anticancer chemotherapeutic agent is selected from teniposide (teniposide)

cisplatin (cisplatin)

satraplatin (satraplatin), carboplatin (carboplatin)

etoposide (etoposide)

doxorubicin (doxorubicin)

its any Lipidosome for example pattern Lay (Caelyx) or

cyclophosphamide (cyclophosphamide) 13-cisRA (13-cis-retinoic acid)

ifosfamide (ifosfamide)

gemcitabine (gemcitabine)

irinotecan (irinotecan) vincristine (vincristine)

actinomycin D (dactinomycin)

methotrexate (methotrexate) with any other chemotherapeutics as herein described, the described any other chemotherapeutics of " other chemotherapy " part below for example.In one embodiment of the invention, the dosage range of any anti-IGF1R antibody shown in this article is about 0.3-20mg/kg body weight or is about 40-1200mg/m ².In one embodiment of the invention, IGF1R inhibitor and other anticancer therapy medicine give simultaneously.In one embodiment of the invention, IGF1R inhibitor and other anticancer therapy medicine are non-gives simultaneously.In one embodiment of the invention, antibody comprises IgG constant region.In one embodiment of the invention, curee is people (for example child).In one embodiment of the invention, IGF1R inhibitor is combined and is given with anti-cancer therapies.In one embodiment of the invention, anti-cancer therapies is surgery tumorectomy and/or anticancer radiotherapy.

Accompanying drawing summary

Fig. 1. the IGF1 RmRNA in constitutional tumor of head and neck sample expresses, and uses Taqman to measure.Each point represents the normalization expression of IGF1R in single tissue sample.I, II, III or the IV for stadium of incidence cancer that therefrom obtains the patient of each sample represents.Corresponding to the value of normal structure sample, with " normally ", represent, corresponding to being positioned near tumor cell but do not there is " NAT " expression for value of tissue normal characteristic normal near.

Fig. 2. the western blot analysis (upper figure) of the protein expression level of total IGF1R, IGF-I and IGF-II in each cell line.By each cell line, be secreted into IGF1 around of growth medium and amount (ng IGF1 or IGF2 albumen/10 of IGF2 ⁶individual cell) (figure below).

Fig. 3 (a)-3 (c). the inhibition of the anti-IGF1R antibody LCF/HCA of variable concentrations to squamous cell carcinoma in body outer cell line.(a): cell line SCC 15; (b): cell line SCC 25; (c): cell line SCC 9.

Fig. 4. with respect to the expression of IGF2 in normal structure Primary Gastric adenocarcinoma tumor tissues.

Fig. 5. the expression of different proteins in some gastric cancer and ovarian cancer cell line, with western blot analysis, measure.

Fig. 6. the anti-IGF1R antibody LCF/HCA of variable concentrations suppresses the growth in vitro of squamous cell cancerous cell (cell line SNU-16).

Fig. 7. be exposed to after anti-IGF1R antibody LCF/HCA or contrast immunoglobulin time dependent renal cell carcinoma tumor growth in mice xenograft models.

Fig. 8. when being exposed to the anti-IGF1R antibody LCF/HCA of variable concentrations, the growth in vitro of melanoma cell series A375-SM suppresses.

Fig. 9. when being exposed to anti-IGF1R antibody LCF/HCA, in mice, the tumor growth of squamous cell cancerous cell line SCC15 suppresses.

In Figure 10 .EW5 (Ewing sarcoma), OS-1 (osteosarcoma) and OS-9 (osteosarcoma) xenograft tumor model to the time dependent evaluation of gross tumor volume.

Detailed Description Of The Invention

The present invention includes the compositions and the method that are used for the treatment of or prevent to comprise following non-cancer medical conditions and cancer: incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma, chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, Burkitt lymphoma, mycosis fungoides, seary's syndrome, cutaneous T cell lymphoma, chronic myeloproliferative diseases) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor, hepatocarcinoma, gastric cancer, gigantism, pituitary adenoma, psoriasis and kidney rhabdoid tumor.Can for example, by giving IGF1R inhibitor (anti-IGF1R antibody) treatment or prophylaxis of cancer.Antibody can with other chemotherapeutics coupling, for example any anticancer chemotherapeutic agent in all anticancer chemotherapeutic agent as described herein.

Term IGF1R, IGFR1 and IGF-1R are synonyms, all represent type-1 insulin like growth factor receptor.

IGF1R inhibitor

Term " IGF1R inhibitor " or " IGF1R antagonist " etc. comprise reducing expresses, ligand binding (being for example combined with IGF-1 and/or IGF-2), any material of any other biological activity of kinase activity (for example autophosphorylation active) or IGF1R (for example mediating anchorage-independent cell growth) and phosphoric acid-IRS-1 level, it can cause (the research worker for example by implementer, doctor or veterinary) definite tissue, system, curee or patient's any biologically or medical response, described reaction comprises any measurable sign that alleviates non-cancer medical conditions and cancer (for example tumor growth) in any degree, symptom and/or clinical marker and/or prevention, slow down or stop process and the transfer of described cancer, for example incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma, chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, Burkitt lymphoma, mycosis fungoides, seary's syndrome, cutaneous T cell lymphoma, chronic myeloproliferative diseases) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor, hepatocarcinoma, gastric cancer, gigantism, pituitary adenoma, psoriasis and kidney rhabdoid tumor.

In one embodiment of the invention, the IGF1R inhibitor that gives patient according to method of the present invention is and the separation antibody of IGF-1R (for example people IGF1R) specific binding or its Fab (for example monoclonal antibody (for example completely human monoclonal antibodies), polyclonal antibody, bi-specific antibody, Fab antibody fragment, F (ab) ₂antibody fragment, Fv antibody fragment (for example VH or VL), single-chain Fv antibody fragment, dsFv antibody fragment, humanized antibody, chimeric antibody or anti-idiotype antibody), such as Burtrum etc., CancerResearch 63:8912-8921 (2003); Disclosed any antibody or antibody fragment in french patent application FR2834990, FR2834991 and FR2834900 and PCT application publication number WO 03/100008, WO 03/59951, WO2006/13472, WO 04/71529, WO 03/106621, WO 04/83248, WO 04/87756, WO 05/16970 and WO 02/53596.

In one embodiment of the invention, the IGF1R inhibitor that the method according to this invention gives patient is separated glucagon like growth factor-1 receptor (IGF1R) antibody, and described antibody comprises ripe 19D12/15H12 light chain C, light chain D, light chain E or light chain F (LCC, LCD, LCE or LCF) and ripe 19D12/15H12 heavy chain A or heavy chain B (HCA or HCB).In one embodiment of the invention, antibody comprises ripe LCF and ripe HCA (LCF/HCA).In one embodiment of the invention, the IGF1R inhibitor that gives patient according to the inventive method is the separation antibody with IGF1R specific binding, its one or more complementary determining regions (CDR) that comprise 19D12/15H12 light chain C, light chain D, light chain E or light chain F and/or 19D12/15H12 heavy chain A or heavy chain B (for example all 3 light chain CDR and all 3 heavy chain CDR).

Aminoacid sequence and the nucleotide sequence of some antibody chain of the present invention see below.Type with dotted line underscore represents signal peptide.Type with solid line underscore represents CDR.General type represents framework region.Ripe fragment lacks signal peptide.

The 19D12/15H12 light chain C (SEQ ID NO:1) modifying

GAA ATT GTG CTG ACT CAG AGC CCA GAC TCT CTG TCT GTG ACT CCA

GGC GAG AGA GTC ACC ATC ACC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC

TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG TCT CCA AAG CTT CTC ATC AAG

TAT GCA TCC CAG TCC CTC TCA GGG GTC CCC TCG AGG TTC AGT GGC AGT GGA

TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGC CTC GAG GCT GAA GAT GCT

GCA GCG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA

GGG ACC AAG GTG GAG ATC AAA CGT ACG

(SEQ ID NO：2)

E I V L T Q S P D S L S V T P

G E R V T I T C R A S Q S I G S S

L H W Y Q Q K P G Q S P K L L I K

Y A S Q S L S G V P S R F S G S G

S G T D F T L T I S S L E A E D A

A A Y Y C H Q S S R L P H T F G Q

G T K V E I K R T

The 19D12/15H12 light chain D (SEQ ID NO:3) modifying

GAA ATT GTG CTG ACT CAG AGC CCA GAC TCT CTG TCT GTG ACT CCA

GGC GAG AGA GTC ACC ATC ACC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC

TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG TCT CCA AAG CTT CTC ATC AAG

TAT GCA TCC CAG TCC CTC TCA GGG GTC CCC TCG AGG TTC AGT GGC AGT GGA

TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGC CTC GAG GCT GAA GAT TTC

GCA GTG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA

GGG ACC AAG GTG GAG ATC AAA CGT ACG

(SEQ ID NO：4)

E I V L T Q S P D S L S V T P

G E R V T I T C R A S Q S I G S S

L H W Y Q Q K P G Q S P K L L I K

Y A S Q S L S G V P S R F S G S G

S G T D F T L T I S S L E A E D F

A V Y Y C H Q S S R L P H T F G Q

G T K V E I K R T

The 19D12/15H12 light chain E (SEQ ID NO:5) modifying

GAA ATT GTG CTG ACT CAG AGC CCA GGT ACC CTG TCT GTG TCT CCA

GGC GAG AGA GCC ACC CTC TCC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC

TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG GCT CCA AGG CTT CTC ATC AAG

TAT GCA TCC CAG TCC CTC TCA GGG ATC CCC GAT AGG TTC AGT GGC AGT GGA

TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGA CTG GAG CCT GAA GAT GCT

GCA GCG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA

GGG ACC AAG GTG GAG ATC AAA CGT ACA

(SEQ ID NO：6)

E I V L T Q S P G T L S V S P

G E R A T L S C R A S Q S I G S S

L H W Y Q Q K P G Q A P R L L I K

Y A S Q S L S G I P D R F S G S G

S G T D F T L T I S R L E P E D A

A A Y Y C H Q S S R L P H T F G Q

G T K V E I K R T

The 19D12/15H12 light chain F (SEQ ID NO:7) modifying

GAA ATT GTG CTG ACT CAG AGC CCA GGT ACC CTG TCT GTG TCT CCA

GGC GAG AGA GCC ACC CTC TCC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC

TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG GCT CCA AGG CTT CTC ATC AAG

TAT GCA TCC CAG TCC CTC TCA GGG ATC CCC GAT AGG TTC AGT GGC AGT GGA

TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGA CTG GAG CCT GAA GAT TTC

GCA GTG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA

GGG ACC AAG GTG GAG ATC AAA CGT ACA

(SEQ ID NO：8)

E I V L T Q S P G T L S V S P

G E R A T L S C R A S Q S I G S S

L H W Y Q Q K P G Q A P R L L I K

Y A S Q S L S G I P D R F S G S G

S G T D F T L T I S R L E P E D F

A V Y Y C H Q S S R L P H T F G Q

G T K V E I K R T

The 19D12/15H12 heavy chain A (SEQ ID NO:9) modifying

GAG GTT CAG CTG GTG CAG TCT GGG GGA GGC TTG GTA AAG CCT GGG

GGG TCC CTG AGA CTC TCC TGT GCA GCC TCT GGA TTC ACC TTC AGT AGC TTT

GCT ATG CAC TGG GTT CGC CAG GCT CCA GGA AAA GGT CTG GAG TGG ATA TCA

GTT ATT GAT ACT CGT GGT GCC ACA TAC TAT GCA GAC TCC GTG AAG GGC CGA

TTC ACC ATC TCC AGA GAC AAT GCC AAG AAC TCC TTG TAT CTT CAA ATG AAC

AGC CTG AGA GCC GAG GAC ACT GCT GTG TAT TAC TGT GCA AGA CTG GGG AAC

TTC TAC TAC GGT ATG GAC GTC TGG GGC CAA GGG ACC ACG GTC ACC GTC TCC

TCA

(SEQ ID NO：10)

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ser

Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg

Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn

Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Leu Gly Asn

Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser

Ser

The 19D12/15H12 heavy chain B (SEQ ID NO:11) modifying

GAG GTT CAG CTG GTG CAG TCT GGG GGA GGC TTG GTA CAG CCC GGG

GGG TCC CTG AGA CTC TCC TGT GCA GCC TCT GGA TTC ACC TTC AGT AGC TTT

GCT ATG CAC TGG GTT CGC CAG GCT CCA GGA AAA GGT CTG GAG TGG ATA TCA

GTT ATT GAT ACT CGT GGT GCC ACA TAC TAT GCA GAC TCC GTG AAG GGC CGA

TTC ACC ATC TCC AGA GAC AAT GCC AAG AAC TCC TTG TAT CTT CAA ATG AAC

AGC CTG AGA GCC GAG GAC ACT GCT GTG TAT TAC TGT GCA AGA CTG GGG AAC

TTC TAC TAC GGT ATG GAC GTC TGG GGC CAA GGG ACC ACG GTC ACC GTC TCC

TCA

(SEQ ID NO：12)

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ser

Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg

Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn

Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Leu Gly Asn

Phe Tyr Tyr Gly Met Asp ValTrp Gly Gln Gly Thr Thr Val Thr Val Ser

Ser

The present invention includes wherein antibody chain described herein is guarded and replaces but the embodiment of not appreciable impact antibody binding activity by one or more sudden changes.

The plasmid that comprises the CMV promoter being effectively connected with heavy chain with 15H12/19D12 light chain has been deposited in American type culture collection (American TypeCulture Collection, ATCC) on May 21st, 2003; 10801University Boulevard; Manassas, Virginia 20110-2209.Preservation title and the ATCC preserving number of plasmid are as follows:

(i) CMV promoter-15H12/19D12HCA (γ 4)-

Preservation title: " 15H12/19D12HCA (γ 4) ";

ATCC preserving number: PTA-5214;

(ii) CMV promoter-15H12/19D12HCB (γ 4)-

Preservation title: " 15H12/19D12HCB (γ 4) ";

ATCC preserving number: PTA-5215;

(iii) CMV promoter-15H12/19D12HCA (γ 1)-

Preservation title: " 15H12/19D12HCA (γ 1) ";

ATCC preserving number: PTA-5216;

(iv) CMV promoter-15H12/19D12LCC (κ)-

Preservation title: " 15H12/19D12LCC (κ) ";

ATCC preserving number: PTA-5217;

(v) CMV promoter-15H12/19D12LCD (κ)-

Preservation title: " 15H12/19D12LCD (κ) ";

ATCC preserving number: PTA-5218;

(vi) CMV promoter-15H12/19D12LCE (κ)-

Preservation title: " 15H12/19D12LCE (κ) ";

ATCC preserving number: PTA-5219; With

(vii) CMV promoter-15H12/19D12LCF (κ)-

Preservation title: " 15H12/19D12LCF (κ) ";

ATCC preserving number: PTA-5220;

The present invention includes the method and composition (for example method and composition disclosed herein) that comprises anti-IGF1R antibody and Fab thereof, described anti-IGF1R antibody and Fab thereof comprise light chain immunoglobulin and/or heavy chain immunoglobulin or its ripe fragment being positioned on any aforementioned plasmid that is preserved in ATCC.

In one embodiment of the invention, with the antibody of people IGF1R " specificity " combination, in conjunction with Kd be about 10 ^-8m or 10 ^-7m or more fractional value; Or, in one embodiment of the invention, in conjunction with Kd be about 1.28 * 10 ^-10m or more fractional value (Biacore algoscopy), or the Kd of combination is about 2.05 * 10 ^-12m or lower numerical value (KinExA algoscopy).In another embodiment, be only combined with people IGF1R by any ormal weight with the antibody of people IGF1R " specificity " combination, and not with other protein binding.

In one embodiment of the invention, according to the inventive method, give patient's IGF1R inhibitor packages containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2002/53596, this application is all attached to herein by reference.

For example, in one embodiment of the invention, antibody comprises variable region of light chain and/or variable region of heavy chain, this variable region of light chain comprises the SEQ ID NO:2 being selected from shown in WO 2002/53596,6,10,14,18,22,47 and 51 aminoacid sequence, and this variable region of heavy chain comprises the SEQ ID NO:4 being selected from shown in WO 2002/53596,8,12,16,20,24,45 and 49 aminoacid sequence.In one embodiment of the invention, antibody comprises antibody 2.12.1, the 2.13.2,2.14.3,3.1.1,4.9.2 and the 4.17.3 that are selected from WO 2002/53596 heavy chain and/or light chain.

In one embodiment of the invention, the IGF1R inhibitor packages that can give patient according to the inventive method is containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2003/59951, and this application is all attached to herein by reference.For example, in one embodiment of the invention, antibody comprises variable region of light chain and/or variable region of heavy chain, this variable region of light chain comprises the SEQ IDNO:54 shown in WO 2003/59951,61 and 65 aminoacid sequence, and this variable region of heavy chain comprises the SEQ ID NO:69 being selected from shown in WO2003/59951,75,79 and 83 aminoacid sequence.

In one embodiment of the invention, the IGF1R inhibitor packages that can give patient according to the inventive method is containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2004/83248, and this application is all attached to herein by reference.For example, in one embodiment of the invention, antibody comprises variable region of light chain and/or variable region of heavy chain, this variable region of light chain comprises the SEQID NO:109 being selected from shown in WO 2004/83248, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141 and 143 aminoacid sequence, this variable region of heavy chain comprises the SEQ ID NO:108 being selected from shown in WO 2004/83248, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140 and 142 aminoacid sequence.In one embodiment of the invention, antibody comprises light chain and/or heavy chain, and this light chain and/or heavy chain are selected from light chain and/or the heavy chain of PINT-6A1, PINT-7A2, PINT-7A4, PINT-7A5, PINT-7A6, PINT-8A1, PINT-9A2, PINT-11A1, PINT-11A2, PINT-11A3, PINT-11A4, PINT-11A5, PINT-11A7, PINT-12A1, PINT-12A2, PINT-12A3, PINT-12A4 and PINT-12A5 in WO 2004/83248.

In one embodiment of the invention, the IGF1R inhibitor packages that can give patient according to the inventive method is containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2003/106621, and this application is all attached to herein by reference.For example, in one embodiment of the invention, antibody comprises variable region of light chain and/or variable region of heavy chain, this variable region of light chain comprises the SEQ ID NO:8-12 that is selected from shown in WO 2003/106621,58-69,82-86,90,94,96,98 aminoacid sequence, and this variable region of heavy chain comprises SEQ ID NO:7,13, the 70-81,87,88 being selected from shown in WO 2003/106621,92 aminoacid sequence.

In one embodiment of the invention, the IGF1R inhibitor packages that can give patient according to the inventive method is containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2004/87756, and this application is all attached to herein by reference.For example, in one embodiment of the invention, antibody comprises variable region of light chain or variable region of heavy chain, the aminoacid sequence that this variable region of light chain comprises the SEQ ID NO:2 shown in WO 2004/87756, the aminoacid sequence that this variable region of heavy chain comprises the SEQ IDNO:1 shown in WO 2004/87756.

In one embodiment of the invention, the IGF1R inhibitor packages that can give patient according to the inventive method is containing any light chain immunoglobulin and/or the heavy chain immunoglobulin shown in published international application no WO 2005/16970, and this application is all attached to herein by reference.For example, in one embodiment of the invention, antibody comprises variable region of light chain and/or variable region of heavy chain, this variable region of light chain comprises the SEQ IDNO:6 shown in WO 2005/16970 or 10 aminoacid sequence, the aminoacid sequence that this variable region of heavy chain comprises the SEQ ID NO:2 shown in WO 2005/16970.

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise immunoglobulin heavy chain variable region, and it comprises and is selected from following aminoacid sequence:

1 grlgqawrsl rlscaasgft fsdyymswir qapgkglewv syisssgstr

51 dyadsvkgrf tisrdnakns lylqmnslra edtavyycvr dgvettfyyy

101 yygmdvwgqg ttvtvssast kgpsvfplap csrstsesta algclvkdyf

151 pepvtvswns galtsgvhtf psca

(SEQ ID NO：13)

1 vqllesgggl vqpggslrls ctasgftfss yamnwvrqap gkglewvsai

51 sgsggttfya dsvkgrftis rdnsrttlyl qmnslraedt avyycakdlg

101 wsdsyyyyyg mdvwgqgttv tvss

(SEQ ID NO：14)

1 gpglvkpset lsltctvsgg sisnyywswi rqpagkglew igriytsgsp

51 nynpslksrv tmsvdtsknq fslklnavta adtavyycav tifgvviifd

101 ywgqgtlvtv ss

(SEQ ID NO：15)

1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsa

51 isgsggityy adsvkgrfti srdnskntly lqmnslraed tavyycakdl

101 gygdfyyyyy gmdvwgqgtt vtvss

(SEQ ID NO：16)

1 pglvkpsetl sltctvsggs issyywswir qppgkglewi gyiyysgstn

51 ynpslksrvt isvdtsknqf slklssvtaa dtavyycart ysssfyyygm

101 dvwgqgttvt vss

(SEQ ID NO：17)

1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsg

51 itgsggstyy adsvkgrfti srdnskntly lqmnslraed tavyycakdp

101 gttvimswfd pwgqgtlvtv ss

(SEQ ID NO：18)

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise immunoglobulin light chain variable region, and it comprises and is selected from following aminoacid sequence:

1 asvgdrvtft crasqdirrd lgwyqqkpgk apkrliyaas rlqsgvpsrf

51 sgsgsgteft ltisslqped fatyyclqhn nyprtfgqgt eveiirtvaa

101 psvfifppsd eqlksgtasv vcllnnfypr eakvqw

(SEQ ID NO：19)

1 diqmtqfpss lsasvgdrvt itcrasqgir ndlgwyqqkp gkapkrliya

51 asrlhrgvps rfsgsgsgte ftltisslqp edfatyyclq hnsypcsfgq

101 gtkleik

(SEQ ID NO：20)

1 sslsasvgdr vtftcrasqd irrdlgwyqq kpgkapkrli yaasrlqsgv

51 psrfsgsgsg teftltissl qpedfatyyc lqhnnyprtf gqgteveiir

(SEQ ID NO：21)

1 diqmtqspss lsasvgdrvt itcrasqgir sdlgwfqqkp gkapkrliya

51 asklhrgvps rfsgsgsgte ftltisrlqp edfatyyclq hnsypltfgg

101 gtkveik

(SEQ ID NO：22)

1 gdrvtitcra sqsistflnw yqqkpgkapk llihvasslq ggvpsrfsgs

51 gsgtdftlti sslqpedfat yycqqsynap ltfgggtkve ik

(SEQ ID NO：23)

1 ratlscrasq svrgrylawy qqkpgqaprl liygassrat gipdrfsgsg

51 sgtdftltis rlepedfavf ycqqygsspr tfgqgtkvei k

(SEQ ID NO：24)

In one embodiment of the invention, anti-IGF1R antibody comprises light chain immunoglobulin or its ripe fragment (lacking signal sequence) or its variable region, and it comprises following aminoacid sequence:

1 mdmrvpaqll gllllwfpga rc diqmtqsp sslsasvgdr vtitcrasqg

51 irndlgwyqq kpgkapkrli yaasslqsgv psrfsgsgsg teftltissl

101 qpedfatyyc lqhnsypwtf gqgtkveikr tvaapsvfif ppsdeqlksg

151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst

201 ltlskadyek hkvyacevth qglsspvtks fnrgec ；

(SEQ ID NO：25)

1 mdmrvpaqll gllllwfpga rc diqmtqsp sslsasvgdr vtftcrasqd

51 irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg teftltissl

101 qpedfatyyc lqhnnyprtf gqgteveiir tvaapsvfif ppsdeqlksg

151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst

201 ltlskadyek hkvyacevth qglsspvtks fnrgec ；

(SEQ ID NO：26)

1 mdmrvpaqll gllllwfpga rc diqmtqsp sslsasvgdr vtitcrasqg

51 irndlgwyqq kpgkapkrli yaasslqsgv psrfsgsgsg teftltissl

101 qpedfatyyc lqhnsypytf gqgtkleikr tvaapsvfif ppsdeqlksg

151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst

201 ltlskadyek hkvyacevth qglsspvtks fnrgec ；

(SEQ ID NO：27)

Or

1 mdmrvpaqll gllllwfpga rc diqmtqfp sslsasvgdr vtitcrasqg

51 irndlgwyqq kpgkapkrli yaasrlhrgv psrfsgsgsg teftltissl

101 qpedfatyyc lqhnsypcsf gqgtkleikr tvaapsvfif ppsdeqlksg

151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst

201 ltlskadyek hkvyacevth qglsspvtks fnrgec

(SEQ ID NO：28)。

In one embodiment of the invention, signal sequence is amino acid/11-22 of SEQ ID NO:25-28.In one embodiment of the invention, there is underscore ripe variable region.In one embodiment of the invention, CDR is boldface letter/italics.In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise one or more CDR as implied above (for example 3 light chain CDR).

In one embodiment of the invention, anti-IGF1R antibody comprises heavy chain immunoglobulin or its ripe fragment (lacking signal sequence) or its variable region, and it comprises following aminoacid sequence:

1 mefglswvfl vaiikgvqc q vqlvesgggl vkpggslrls caasgftfsd

51 yymswirqap gkglewvsyi sssgstiyya dsvkgrftis rdnaknslyl

101 qmnslraedt avyycarvlr flewllyyyy yygmdvwgqg ttvtvssast

151 kgpsvfplap csrstsesta algclvkdyf pepvtvswns galtsgvhtf

201 pavlqssgly slssvvtvps snfgtqtytc nvdhkpsntk vdktverkcc

251 vecppcpapp vagpsvflfp pkpkdtlmis rtpevtcvvv dvshedpevq

301 fnwyvdgvev hnaktkpree qfnstfrvvs vltvvhqdwl ngkeykckvs

351 nkglpapiek tisktkgqpr epqvytlpps reemtknqvs ltclvkgfyp

401 sdiavewesn gqpennyktt ppmldsdgsf flyskltvdk srwqqgnvfs

451 csvmhealhn hytqkslsls pgk ；

(SEQ ID NO：29)

1 mefglswvfl vaiikgvqc q aqlvesgggl vkpggslrls caasgftfsd

51 yymswirqap gkglewvsyi sssgstrdya dsvkgrftis rdnaknslyl

101 qmnslraedt avyycvrdgv ettfyyyyyg mdvwgqgttv tvssastkgp

151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav

201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec

251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw

301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg

351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi

401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv

451 mhealhnhyt qkslslspgk ；

(SEQ ID NO：30)

1 mefglswlfl vailkgvqc e vqllesgggl vqpggslrls caasgftfss

51 yamswvrqap gkglewvsai sgsggstyya dsvkgrftis rdnskntlyl

101 qmnslraedt avyycakgys sgwyyyyyyg mdvwgqgttv tvssastkgp

151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav

201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec

251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw

301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg

351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi

401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv

451 mhealhnhyt qkslslspgk ；

(SEQ ID NO：31)

Or

1 mefglswlfl vailkgvqc e vqllesgggl vqpggslrls ctasgftfss

51 yamnwvrqap gkglewvsai sgsggttfya dsvkgrftis rdnsrttlyl

101 qmnslraedt avyycakdlg wsdsyyyyyg mdvwgqgttv tvssastkgp

151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav

201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec

251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw

301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg

351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi

401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv

451 mhealhnhyt qkslslspgk

(SEQ ID NO：32)。

In one embodiment of the invention, signal sequence is amino acid/11-19 of SEQ ID NO:29-32.In one embodiment of the invention, there is underscore ripe variable region.In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise one or more CDR as implied above (for example 3 light chain CDR).

In one embodiment of the invention, anti-IGF1R antibody comprises respectively variable region of light chain and variable region of heavy chain pairing, this variable region of light chain comprises in SEQ ID NO:19-24 the aminoacid sequence of any, and this variable region of heavy chain comprises in SEQ ID NO:13-18 the aminoacid sequence of any.In one embodiment of the invention, anti-IGF1R antibody comprises ripe variable region of light chain and variable region of heavy chain pairing, this maturation variable region of light chain comprises in SEQ ID NO:25 or 26 aminoacid sequence of any, and this variable region of heavy chain comprises in SEQ ID NO:29 or 30 aminoacid sequence of any.In one embodiment of the invention, anti-IGF1R antibody comprises ripe variable region of light chain and variable region of heavy chain pairing, this maturation variable region of light chain comprises in SEQ IDNO:27 or 28 aminoacid sequence of any, and this variable region of heavy chain comprises in SEQ ID NO:31 or 32 aminoacid sequence of any.

In one embodiment of the invention, (in one embodiment of the invention, targeting sequencing has underscore for the heavy chain immunoglobulin that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx or ripe fragment or variable region (SEQ IDNO:33); In one embodiment of the invention, CDR is boldface letter/italics):

1 mefglswvfl vaiikgvqcq vqlvesgggl vkpggslrls caasgftfsd

51 yymswirqap gkglewvsyi sssgstrdya dsvkgrftis rdnaknslyl

101 qmnslraedt avyycardgv ettfyyyyyg mdvwgqgttv tvssastkgp

151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav

201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec

251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw

301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg

351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi

401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv

451 mhealhnhyt qkslslspgk

In one embodiment of the invention, the aminoacid 20-470 (SEQ ID NO:33) that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx.

In one embodiment of the invention, maturation immunity globulin variable region of heavy chain (the aminoacid 20-144 of SEQ ID NO:33 that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx; SEQ ID NO:34):

q vqlvesgggl vkpggslrls caasgftfsd yymswirqap gkglswvsyi sssgstrdya

dsvkgrftis rdnaknslyl qmnslraedt avyycardgv ettfyyyyyg mdvwgqgttv tvss

In one embodiment of the invention, (in one embodiment of the invention, targeting sequencing has underscore for the light chain immunoglobulin that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx or ripe fragment or variable region (SEQ IDNO:35); In one embodiment of the invention, CDR is boldface letter/italics):

1 mdmrvpaqll qllllwfpga rcdiqmtqsp sslsasvgdr vtitcrasqd

51 irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg teftltissl

101 qpedfatyyc lqhnnyprtf gqgtkveikr tvaapsvfif ppsdeqlksg

151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst

201 ltlskadyek hkvyacevth qglsspvtks fnrgec

In one embodiment of the invention, the aminoacid 23-236 (SEQ ID NO:35) that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx.

In one embodiment of the invention, maturation immunity globulin variable region of light chain (the aminoacid 23-130 of SEQ ID NO:35 that anti-IGF1R antibody of the present invention or its Fab comprise 2.12.1fx; SEQ ID NO:36):

diqmtqsp sslsasvgdr vtitcrasqd irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg

teftltissl qpedfatyyc lqhnnyprtf gqgtkveikr

In one embodiment of the invention, anti-IGF1R antibody or its Fab comprise light chain immunoglobulin chain and heavy chain immunoglobulin chain or are comprised of light chain immunoglobulin chain and heavy chain immunoglobulin chain, the aminoacid 23-236 that this light chain immunoglobulin chain comprises 2.12.1fx (SEQ ID NO:35) or formed by the aminoacid 23-236 (SEQ ID NO:35) of 2.12.1fx, the aminoacid 20-470 that this heavy chain immunoglobulin chain comprises 2.12.1fx (SEQ IDNO:33) or formed by the aminoacid 20-470 (SEQ ID NO:33) of 2.12.1fx.

In one embodiment of the invention, anti-IGF1R antibody or its Fab comprise one or more 2.12.1fxCDR as implied above (for example 3 light chain CDR and/or 3 heavy chain CDR).

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab or its Fab comprise humanization 7C10 immunoglobulin light chain variable region; Form 1 (SEQ ID NO:37):

1 dvvmtqspls lpvtpgepas iscrssqsiv hsngntylqw ylqkpgqspq

51 lliykvsnrl ygvpdrfsgs gsgtdftlki srveaedvgv yycfqgshvp

101 wtfgqgtkve ik

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise humanization 7C10 immunoglobulin light chain variable region; Form 2 (SEQ ID NO:38):

1 divmtqspls lpvtpgepas iscrssqsiv hsngntylqw ylqkpgqspq

51 lliykvsnrl ygvpdrfsgs gsgtdftlki srveaedvgv yycfqgshvp

101 wtfgqgtkve ik

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise humanization 7C10 immunoglobulin heavy chain variable region; Form 1 (SEQ ID NO:39):

1 qvqlqesgpg lvkpsetlsl tctvsgysit ggylwnwirq ppgkglewmg

51 yisydgtnny kpslkdriti srdtsknqfs lklssvtaad tavyycaryg

101 rvffdywgqg tlvtvss

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise humanization 7C10 immunoglobulin heavy chain variable region; Form 2 (SEQ ID NO:40):

1 qvqlqesgpg lvkpsetlsl tctvsgysit ggylwnwirq ppgkglewig

51 yisydgtnny kpslkdrvti srdtsknqfs lklssvtaad tavyycaryg

101 rvffdywgqg tlvtvss

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise humanization 7C10 immunoglobulin heavy chain variable region; Form 3 (SEQ ID NO:41):

1 qvqlqesgpg lvkpsetlsl tctvsgysis ggylwnwirq ppgkglewig

51 yisydgtnny kpslkdrvti svdtsknqfs lklssvtaad tavyycaryg

101 rvffdywgqg tlvtvss

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise A12 immunoglobulin heavy chain variable region (SEQ ID NO:42):

1 evqlvqsgae vkkpgssvkv sckasggtfs syaiswvrqa pgqglewmgg

51 iipifgtany aqkfqgrvti tadkststay melsslrsed tavyycarap

101 lrflewstqd hyyyyymdvw gkgttvtvss

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise A12 immunoglobulin light chain variable region (SEQ ID NO:43):

1 sseltqdpav svalgqtvri tcqgdslrsy yaswyqqkpg qapvlviygk

51 nnrpsgipdr fsgsssgnta sltitgaqae deadyycnsr dnsdnrllfg

101 ggtkltvls

Or

(SEQ ID NO：105)：

1 sseltqdpav svalgqtvri tcqgdslrsy yatwyqqkpg qapilviyge

51 nkrpsgipdr fsgsssgnta sltitgaqae deadyycksr dgsgqhlvfg

101 ggtkltvlg

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise IA immunoglobulin heavy chain variable region (SEQ ID NO:44):

1 evqlvqsggg lvhpggslrl scagsgftfr nyamywvrqa pgkglewvsa

51 igsgggtyya dsvkgrftis rdnaknslyl qmnslraedm avyycarapn

101 wgsdafdiwg qgtmvtvss ；

Optionally comprise one or more following sudden changes: R30, S30, N31, S31, Y94, H94, D104, E104.

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise 1A immunoglobulin light chain variable region (SEQ ID NO:45):

1 diqmtqspss lsasvgdrvt itcrasqgis swlawyqqkp ekapksliya

51 asslqsgvps rfsgsgsgtd ftltisslqp edfatyycqq ynsypptfgp

101 gtkvdik ；

Optionally comprise one or more following sudden changes: P96, I96, P100, Q100, R103, K103, V104, L104, D105, E105.

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 8A1 (SEQ ID NO:46):

1 evqlvqsgae vkkpgeslti sckgpgynff nywigwvrqm pgkglewmgi

51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi

101 rycpggrcys gyygmdvwgq gtmvtvssgg ggsggggsgg ggsseltqdp

151 avsvalgqtv ritcqgdslr syyaswyqqk pgqapvlviy gknnrpsgip

201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhvv fgggtkltvl

251 g

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 9A2 (SEQ ID NO:47):

1 qvqlvqsgae vrkpgasvkv scktsgytfr nydinwvrqa pgqglewmgr

51 isghygntdh aqkfqgrftm tkdtststay melrsltfdd tavyycarsq

101 wnvdywgrgt lvtvssgggg sggggsgggg salnfmltqp hsvsespgkt

151 vtisctrssg siasnyvqwy qqrpgssptt vifednrrps gvpdrfsgsi

201 dtssnsaslt isglktedea dyycqsfdst nlvvfgggtk vtvlg

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 11A4 (SEQ ID NO:48):

1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsa

51 isgsggstyy adsvkgrfti srdnskntly lqmnslraed tavyycassp

101 yssrwysfdp wgqgtmvtvs sggggsgggg sggggsalsy eltqppsvsv

151 spgqtatitc sgddlgnkyv swyqqkpgqs pvlviyqdtk rpsgiperfs

201 gsnsgniatl tisgtqavde adyycqvwdt gtvvfgggtk ltvlg

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 7A4 (SEQ ID NO:49):

1 evqlvqsgae vkkpgeslti sckgsgynff nywigwvrqm pgkdlewmgi

51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi

101 rycpggrcys gyygmdvwgq gtmvtvssgg gssggggsgg ggsseltqdp

151 avsvalgqtv ritcrgdslr nyyaswyqqk pgqapvlviy gknnrpsgip

201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhmv fgggtkltvl

251 g

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 11A1 (SEQ ID NO:50):

1 evqlvesggg vvqpgrslrl scaasgftfs dfamhwvrqi pgkglewlsg

51 lrhdgstayy agsvkgrfti srdnsrntvy lqmnslraed tatyycvtgs

101 gssgphafpv wgkgtlvtvs sggggsgggg sggggsalsy vltqppsasg

151 tpgqrvtisc sgsnsnigty tvnwfqqlpg tapklliysn nqrpsgvpdr

201 fsgsksgtsa slaisglqse deadyycaaw ddslngpvfg ggtkvtvlg

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise single-chain antibody (fv) 7A6 (SEQ ID NO:51):

1 evqlvqsgae vkkpgeslti sckgsgynff nywigwvrqm pgkglewmgi

51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi

101 rycpggrcys gyygmdvwgq gtlvtvssgg ggsggggsgg ggsseltqdp

151 avsvalgqtv ritcqgdslr syytnwfqqk pgqapllvvy aknkrpsgip

201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhvv fgggtkltvl

251 g

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab (for example heavy chain or light chain immunoglobulin) comprise one or more following complementary determining regions (CDR) that are selected from:

sywmh(SEQ ID NO：52)；

einpsngrtnynekfkr(SEQ ID NO：53)；

grpdyygsskwyfdv(SEQ ID NO：54)；

rssqsivhsnvntyle(SEQ ID NO：55)；

Kvsnrfs (SEQ ID NO:56); With

fqgshvppt(SEQ ID NO：57)。

In one embodiment of the invention, anti-IGF1R antibody of the present invention or its Fab comprise and are selected from following heavy chain immunoglobulin variable region:

1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgqgttv tvs

(SEQ ID NO：58)；

1 qvqfqqsgae lvkpgasvkl sckasgytft sylmhwikqr pgrglewigr

51 idpnnvvtkf nekfkskatl tvdkpsstay melssltsed savyycarya

101 ycrpmdywgq gttvtvss

(SEQ ID NO：59)；

1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgagttv tvs

(SEQ ID NO：60)；

1 qvqlqqsgae lmkpgasvki sckatgytfs sfwiewvkqr pghglewige

51 ilpgsggthy nekfkgkatf tadkssntay mqlssltsed savyycargh

101 syyfydgdyw gqgtsvtvss

(SEQ ID NO：61)；

1 qvqlqqpgsv lvrpgasvkl sckasgytft sswihwakqr pgqglewige

51 ihpnsgntny nekfkgkatl tvdtssstay vdlssltsed savyycarwr

101 ygspyyfdyw gqgttltvss

(SEQ ID NO：62)；

1 qvqlqqpgae lvkpgasvkl sckasgytft sywmhwvkqr pgrglewigr

51 idpnsggtky nekfkskatl tvdkpsstay mqlssltsed savyycaryd

101 yygssyfdyw gqgttltvss

(SEQ ID NO：63)；

1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgqgttv tvs

(SEQ ID NO：64)；

1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgagttv tvss

(SEQ ID NO：65)；

1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgqgttv tvss

(SEQ ID NO：66)；

1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgrglewigr

51 idpnsggtky nekfkskatl tvdkpsstay mqlssltsed savyycaryd

101 yygssyfdyw gqgttvtvss

(SEQ ID NO：67)；

1 qiqlqqsgpe lvrpgasvki sckasgytft dyyihwvkqr pgeglewigw

51 iypgsgntky nekfkgkatl tvdtssstay mqlssltsed savyfcargg

101 kfamdywgqg tsvtvss

(SEQ ID NO：68)；

1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige

51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr

101 pdyygsskwy fdvwgagttv tvss

(SEQ ID NO：69)；

1 qiqlqqsgpe lvkpgasvki sckasgytft dyyinwmkqk pgqglewigw

51 idpgsgntky nekfkgkatl tvdtssstay mqlssltsed tavyfcarek

101 ttyyyamdyw gqgtsvtvsa

(SEQ ID NO：70)；

1 vqlqqsgael mkpgasvkis ckasgytfsd ywiewvkqrp ghglewigei

51 lpgsgstnyh erfkgkatft adtssstaym qlnsltseds gvyyclhgny

101 dfdgwgqgtt ltvss

(SEQ ID NO:71); With

1 qvqllesgae lmkpgasvki sckatgytfs sfwiewvkqr pghglewige

51 ilpgsggthy nekfkgkatf tadkssntay mqlssltsed savyycargh

101 syyfydgdyw gqgtsvtvss

(SEQ ID NO：72)；

And/or be selected from following light chain immune globulin variable region:

1 dvlmtqipvs lpvslgdqas iscrssqiiv hnngntylew ylqkpgqspq

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 ftfgsgtkle ikr

(SEQ ID NO：73)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：74)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：75)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：76)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 1liykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：77)；

1 dvlmtqtpls lpvslgdqas iscrssqxiv hsngntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 xtfgggtkle ikr

(SEQ ID NO：78)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：79)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：80)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：81)；

1 dvlmtqipvs lpvslgdqas iscrssqiiv hnngntylew ylqkpgqspq

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 ftfgsgtkle ikr

(SEQ ID NO：82)；

1 dvlmtqtpls lpvslgdqas iscrfsqsiv hsngntylew ylqksgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 rtfgggtkle ikr

(SEQ ID NO：83)；

1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：84)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：85)；

1 elvmtqtpls lpvslgdqas iscrssqtiv hsngdtyldw flqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：86)；

1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：87)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：88)；

1 dvlmtqtpvs lsvslgdqas iscrssqsiv hstgntylew ylqkpgqspk

51 lliykisnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqashap

101 rtfgggtkle ikr

(SEQ ID NO：89)；

1 dvlmtqtpls lpvslgdqas isckssqsiv hssgntyfew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgship

101 ftfgsgtkle ikr

(SEQ ID NO：90)；

1 dieltqtpls lpvslgdqas iscrssqsiv hsngntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 ytfgggtkle ikr

(SEQ ID NO：91)；

1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：92)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：93)；

1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：94)；

1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：95)；

1 dvlmtqtpls lpvslgdqas iscrsnqtil lsdgdtylew ylqkpgqspk

51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp

101 ptfgggtkle ikr

(SEQ ID NO：96)；

1 dvlmtqtpls lpvslgdqas iscrssqtiv hsngntylew ylqkpgqspk

51 lliykvtnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgthap

101 ytfgggtkle ikr

(SEQ ID NO:97); With

1 dvlmtqtpls lpvslgdqas iscrssqsiv hsngntylew ylqkpgqspk

51 lliysissrf sgvpdrfsgs gsgtdftlki srvqaedlgv yycfqgshvp

101 ytfgggtkle ikr

(SEQ ID NO：98)。

Scope of the present invention comprises the method that wherein gives patient's anti-IGFR body-1 (IGF1R) antibody, and wherein antibody variable region is connected with any constant region for immunoglobulin.In one embodiment of the invention, variable region of light chain is connected with κ chain constant region.In one embodiment of the invention, variable region of heavy chain is connected with γ 1, γ 2, γ 3 or γ 4 chain constant regions.In embodiments of the invention, any immune globulin variable region shown in this paper can be connected with aforementioned any constant region.

In addition, scope of the present invention comprises any antibody or the antibody fragment of the framework region that contains any light chain immunoglobulin shown in one or more CDR (3 light chain CDR and/or 3 heavy chain CDR) and/or this paper or heavy chain immunoglobulin, by following any method, identify: Chothia etc., J.Mol.Biol.186:651-663 (1985); Novotny and Haber, Proc.Natl.Acad.Sci.USA 82:4592-4596 (1985) or Kabat, E.A. etc., sequences ofProteins of Immunological Interest, NIH (NationalInstitutes of Health), Bethesda, Md., (1987)).

In one embodiment of the invention, term used herein " monoclonal antibody " refers to the antibody obtaining in the antibody of the basic homogenizing of a group, that is to say that each antibody that this group comprises is identical except may there is the naturally occurring sudden change of a small amount of possibility.As mentioned above, monoclonal antibody can be by the hybridoma method preparation (Kohler etc. (1975) Nature 256:495) being disclosed by people such as Kohler at first used according to the present invention.

In one embodiment of the invention, bispecific or bifunctional antibody are a kind of artificial hybrid antibodies, have two pairs of different heavy chain/light chains and two different binding sites.Bi-specific antibody can produce by the whole bag of tricks (comprising the fusion of hybridoma or the connection of Fab ' fragment).Referring to such as (1990) Clin.Exp.Immunol.79:315-321 such as Songsivilai, Kostelny etc. (1992) J Immunol.148:1547-1553.In addition, bi-specific antibody can be made into " double-chain antibody (diabody) " (Holliger etc. (1993) PNAS USA 90:6444-6448) or makes " Janusins " (Traunecker etc. (1991) EMBO J.10:3655-3659 with (1992) the Int.J.Cancer Suppl.7:51-52 such as Traunecker).

In one embodiment of the invention, term " fully human antibodies " refers to the antibody that only comprises human normal immunoglobulin's protein sequence.Fully human antibodies, if by mice, mouse cell or produce derived from the hybridoma of mouse cell, can contain non-human (for example Mus) sugar chain.Equally, " mouse antibodies " refers to the antibody that only comprises mouse immuning ball protein protein sequence.

The present invention includes " chimeric antibody "---a kind ofly comprise variable region of the present invention and for example, for example, merge with the antibody district (constant region) that derives from other people or inhuman species (mice, horse, rabbit, Canis familiaris L., cattle, chicken) or chimeric antibody.These antibody can regulate expression or the activity of IGF1R in inhuman species.

" scFv " or " sFv " antibody fragment has antibody V _hdistrict and V _ldistrict, wherein these districts are present on wall scroll polypeptide chain.SFv polypeptide generally also comprises V _hdistrict and V _lpeptide linker between district, it can make sFv form the structure of required antigen combination.Can adopt the technology for generation of single-chain antibody described in document (U.S. Patent number 5,476,786,5,132,405 and 4,946,778) to produce anti-IGF1R specific single-chain antibody.The summary of relevant sFv can be referring to Pluckthun, the Pharmacology of Monoclonal Antibodies, the 113rd volume, Rosenburg and Moore chief editor, Springer-Verlag, N.Y., 269-315 page (1994).

In one embodiment of the invention, " the Fv fragment that disulfide bond is stable " and " dsFv " refer to and comprise variable heavy chain (V _h) and variable light chain (V _l) immunoglobulin that is formed by connecting by disulfide bond.

In the scope of the invention, the Fab of antibody also comprises by the F (ab) that for example pepsin enzymatic cutting IgG produces ₂fragment.Fab fragment can be by for example F (ab) ₂with dithiothreitol, DTT or mercaptoethylmaine reduction, produce.Fab fragment is V _l-C _lchain is by disulfide bond and V _h-CH1 chain connects.F (ab) ₂fragment is two Fab fragments that connect by two disulfide bond successively.F (ab) ₂the Fab of molecule partly comprises F between disulfide bond position _cthe part in district.

Fv fragment is V _ldistrict or V _hdistrict.

According to the aminoacid sequence of immunoglobulin heavy chain constant region, immunoglobulin can be divided into different classifications.Immunoglobulin has five large class: IgA, IgD, IgE, IgG and IgM at least, has severally also can further be divided into subclass (isotype), for example IgG-1, IgG-2, IgG-3 and IgG-4 in these; IgA-1 and IgA-2.Any this antibody-like or its Fab discussed herein all fall within the scope of the present invention.

Anti-IGF1R antibody of the present invention and fragment also can be puted together with chemical part.This chemical part can be especially polymer, radionuclide or cytotoxic factor.In one embodiment of the invention, this chemical part is to increase the antibody molecule polymer of the half life in curee's body.Suitable polymer includes but not limited to Polyethylene Glycol (PEG) (for example molecular weight is the PEG of 2kDa, 5kDa, 10kDa, 12kDa, 20kDa, 30kDa or 40kDa), glucosan and poly glycol monomethyl ether (mPEG).The people such as Lee (1999) (Bioconj.Chem.10:973-981) disclose PEG and have puted together single-chain antibody.The people such as Wen (2001) (Bioconj.Chem.12:545-553) disclose antibody are puted together with the PEG that radioactive metal chelating agen (radiometal chelator) (diethylene-triamine pentaacetic acid (DTPA)) is connected.

Antibody of the present invention and antibody fragment also can be puted together with for example following labelling: ⁹⁹tc, ⁹⁰y, ¹¹¹in, ³²p, ¹⁴c, ¹²⁵i, ³h, ¹³¹i, ¹¹c, ¹⁵o, ¹³n, ¹⁸f, ³⁵s, ⁵¹cr, ⁵⁷to, ²²⁶ra, ⁶⁰co, ⁵⁹fe, ⁵⁷se, ¹⁵²eu, ⁶⁷cU, ²¹⁷ci, ²¹¹at, ²¹²pb, ⁴⁷sc, ¹⁰⁹pd, ²³⁴th and ⁴⁰k, ¹⁵⁷gd, ⁵⁵mn, ⁵²tr and ⁵⁶fe.

Antibody of the present invention and antibody fragment also can be puted together with comprising following fluorescent labeling or chemiluminescent labeling: fluorogen for example rare earth chelate compound, fluorescein and derivant thereof, rhodamine and derivant thereof, isothiocyanate, phycoerythrin, phycocyanin, allophycocyanin, phthalic aldehyde, fluorescamine, ¹⁵²eu, dansyl, umbelliferone, luciferin, luminol labelling, different luminol labelling, aromatics acridinium ester labelling (acridinium ester label), imidazoles labelling, acridinium salt (acridimium salt) labelling, oxalate labelling, aequorin labelling, 2,3-dihydro phthalazine diketone, biotin/avidin, spin labeling and stable free radical.

Antibody and antibody fragment also can be puted together with for example following cytotoxic factor: diphtheria toxin, diphtherotoxin, bacillus pyocyaneus (Pseudomonas aeruginosa) exotoxin A chain, ricin A chain, abrin A chain, mould Rhizoma Nelumbinis toxalbumin II A chain, α-broom aspergillin (alpha-sarcin), Aleurites fordii Hemsl. (Aleurites fordii) albumen and compound (for example fatty acid), carnation toxalbumin, phytolacca american (Phytolacca americana) albumen PAPI, PAPII and PAP-S, Fructus Momordicae charantiae mortifier (Momordica charantia inhibitor), curcin, crotin, Saponaria officinalis (Saponaria officinalis) mortifier, mitogellin, restrictocin, phenomycin and enomycin.

Can apply any method that antibody molecule of the present invention and fragment and different piece are puted together known in the art, comprise the Nature144:945 with method described in Publication about Document: Hunter etc. (1962); David etc. (1974) Biochemistry 13:1014; Pain etc. (1981) J.Immunol.Meth.40:219; And Nygren, J., (1982) Histochem.and Cytochem.30:407.For puting together the method for antibody, be that this area is conventional and well-known.

In one embodiment of the invention, IGF1R inhibitor is BMS-577098

or AEW-541

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any pyrimidine derivatives that for example comprises following core texture described in WO 03/,481 33:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any tyrosine kinase inhibitor that for example comprises following core texture described in WO 03/35614:

(for example

).By giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any tyrosine kinase inhibitor that for example comprises following core texture described in WO 03/35615:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any tyrosine kinase inhibitor that for example comprises following core texture described in WO 03/35616:

(for example

).By giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any tyrosine kinase inhibitor that for example comprises following core texture described in WO 03/35619:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor for example, for example, for also suppressing for example many targeting inhibitors of kinases of VEGF-2R, Kit, FLT3 and/or PDGFR, SU-11 248 (Sunitinib malate (sunitinib malate)) or Bay43-9006 (Sorafenib (sorafenib)).By giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any compound that for example comprises following core texture described in WO 03/24967:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any compound that for example comprises following core texture described in WO 04/30625:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any compound that for example comprises following core texture described in WO 04/30627:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any heteroaryl-aryl urea compounds that for example comprises following core texture described in WO 00/35455:

by giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is any peptide described in WO 03/27246.By giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

In one embodiment of the invention, IGF1R inhibitor is

or disclosed any 4-amino-5-phenyl-7-cyclobutyl-pyrrolo-[2,3-d] pyrimidine derivatives in PCT application publication number WO02/92599.By giving these medicines, treat or prevent the method for any medical conditions described herein all to fall within the scope of the present invention.

The generation of antibody

Can apply the antibody that any suitable method induction has the biological characteristics of required inhibition IGF1R.The suitable monoclonal antibody (mAb) of preparing from various mammalian hosts (such as mice, rodent, primates, people etc.).Can be referring to such as (chief editor) BASIC AND CLINICAL IMMUNOLOGY (the 4th edition) such as Stites to preparing the description of the technology of this class monoclonal antibody, Lange Medical Publications, Los Altos, CA and the list of references wherein quoted; Harlow and Lane (1988) ANTIBODIES:A LABORATORYMANUAL CSH Press; Goding (1986) MONOCLONAL ANTIBODIES:PRINCIPLES AND PRACTICE (second edition) Academic Press, New York, NY.Therefore the various technology that, can know by this area research worker obtain monoclonal antibody.Generally make the animal splenocyte immortalization of the antigen immune through meeting the requirements, conventionally by merging with myeloma cell.Referring to Kohler and Milstein (1976) Eur.J.Immunol.6:511-519.The alternative method of immortalization comprises with Epstein-Barr virus, oncogene or retrovirus retrovirus or other method conversion known in the art.Referring to such as Doyle etc. (chief editor, 1994 and regular supplementary issue) CELL AND TISSUE CULTURE:LABORATORY PROCEDURES, John Wiley and Sons, New York, NY.What the colony being produced by single immortalized cell filtered out generation has the antibody of required specificity and affinity to antigen, and can improve the output that is produced monoclonal antibody by this class cell by various technology, comprise and be injected into vertebrate host peritoneal cavity.Or, can by human B cell, filter out DNA library according to general scheme described in the people such as Huse (Huse etc. (1989) Science 246:1275-1281), carry out the DNA sequence of separated coding monoclonal antibody or its binding fragment.

Other suitable technology comprises with phage or similar substrates antagonist library to be selected.Referring to such as Huse etc., Science 246:1275-1281 (1989); And Ward etc., Nature341:544-546 (1989).Can use and be modified or not adorned polypeptide of the present invention and antibody, comprise chimeric antibody or humanized antibody.Conventionally can, by being covalently or non-covalently connected with the material of detectable signal can be provided, to polypeptide and antibody, carry out labelling.Various labellings and conjugation techniques are known, have a large amount of reports in academic documents and patent documentation.As mentioned above, suitable labelling comprises radionuclide, enzyme, substrate, cofactor, inhibitor, fluorescence part, chemiluminescent moiety, magnetic particle etc.The patent of applying this class labelling instructs and comprises U.S. Patent number 3,817,837,3,850,752,3,939,350,3,996,345,4,277,437,4,275,149 and 4,366,241.Equally, preparing recombination immunoglobulin can be referring to (1989) Proc.Nat ' l Acad.Sci.USA86:10029-10033 such as the U.S. Patent number 4,816,567 of Cabilly and Queen; Or preparing transgenic mice can be referring to (1997) NatureGenetics 15:146-156 such as Mendez.The more multi-method that produces chimeric antibody, humanized antibody and people's antibody is well-known in the art, referring to the U.S. Patent number 5 that gives the people such as Queen such as mandate, 530,101, authorize the U.S. Patent number 5 that gives the people such as Winter, 225,539, authorize the U.S. Patent number 4,816 that gives the people such as Boss, 397, described all patents are all attached to herein by reference.

The mammal cell line that can be used as expressing the host of antibody of the present invention is well-known in the art, and comprising can be available from the many immortalized cells system of American type culture collection (ATCC).These especially comprise Chinese hamster ovary (CHO) cell, NSO, SP2 cell, HeLa cell, young hamster kidney (BHK) cell, monkey-kidney cells (COS), human liver cell cancerous cell (for example Hep G2), A549 cell, 3T3 cell, HEK-293 cell and multiple other cell line.Mammalian host cell comprises the cell of following animal: people, mice, rat, Canis familiaris L., monkey, pig, goat, cattle, horse and hamster.The cell line by mensuration with high expression level is selected particularly preferred cell line.Operable other cell is insect cell line, for example Sf9 cell, Amphibian cell, bacterial cell, plant cell and fungal cell.When the recombinant expression carrier of encoding heavy chain or its antigen-binding portion thereof, light chain and/or its antigen-binding portion thereof is imported to mammalian host cell, by being enough to make antibody at host cell inner expression, more preferably, in a period of time in the culture medium that antibody-secreting is grown to host cell, cultivate host cell to produce antibody.

Can adopt standard protein purification process from culture medium, to reclaim antibody.In addition, can apply multiple known technology improves by the antibody of the present invention of producing expression of cell lines the other parts of its acquisition (or by).For example, glutamine synthetase gene expression system (GS system) is the common method that improves under certain conditions expression.Relevant european patent number 0 216 846,0 256 055 and 0 323 997 and European Patent Application No. 89303964.4 in, discussed in whole or in part GS system.

Probably by different cell line or the antibody of expressing, there is different to each other glycosylations in transgenic animal.Yet all antibody by nucleic acid molecule encoding provided herein, or all antibody that comprise aminoacid sequence provided herein are ingredients of the present invention, no matter whether glycosylation of antibody.

Suitable pUC pUC for generation of anti-IGF1R antibody or its Fab can be referring to the U.S. Patent Application No. US2005/0176099 having announced (separately referring to WO2005/47512).

Other chemotherapeutics

The invention provides the method that is used for the treatment of or prevents any medical conditions described herein, the method by treat can acceptable dose or treatment can receiving amount the anti-IGF1R antibody of for example the present invention as herein described or its Fab and other chemotherapeutics (for example anticancer chemotherapeutic agent and/or emesis chemotherapeutics) coupling.For example, other chemotherapeutics comprises erlotinib (erlotinib), Dasatinib (dasatanib), AMN107 (nilotinib), decatanib, Victibix (panitumumab), amrubicin (amrubicin), Ao Gefu monoclonal antibody (oregovomab), Lep-etu (" the liposomal encapsulated paclitaxel of easy-to-use type (paclitaxel) "), 2-Amino-6-methyl-5-(pyridin-4-ylsulfanyl)-3H-quinazolin-4-one (nolatrexed), azd2171, Ba Tabulin (batabulin), method wood monoclonal antibody (ofatumumab) difficult to understand, prick wooden monoclonal antibody (zanolimumab), Ai Te click woods (edotecarin), tetrandrine (tetrandrine), rubitecan (rubitecan), tesmilifene (tesmilifene), Ao Limeisheng (oblimersen), for western wooden monoclonal antibody (ticilimumab), her wooden monoclonal antibody (ipilimumab), gossypol (gossypol), Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide (cilengitide), gefitinib (gimatecan), IL13-PE38QQR, INO 1001, IPdR, KRX-0402, lucanthone (lucanthone), LY 317615, iodine [ ¹³¹i] anti-tenascin monoclonal antibody 81C6 (neuradiab), vitespan, Rta 744, Sdx 102, talampanel (talampanel), atrasentan (atrasentan), Xr 311, everolimus (everolimus), ET-743 (trabectedin), effect of nano-paclitaxel injection suspension (abraxane), TLK 286, AV-299, DN-101, pazopanib (pazopanib), GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD1152, grace is pricked cry loudly woods (enzastaurin), ZD6474 (vandetanib), ARQ-197, MK-0457, MLN8054, PHA-739358, R-763 or AT-9263.

Effect of nano-paclitaxel injection suspension (abraxane) is the injection suspension of the paclitaxel protein binding particle of the paclitaxel that comprises albumin bound form, and mean diameter is about 130 nanometers.The usage of effect of nano-paclitaxel injection suspension is before intravenous infusion, by the extremely yellow aseptic freeze-dried powder 20mL 0.9% sodium chloride injection USP reprovision of white.The bottle that each single is used is equipped with 100mg paclitaxel and about 900mg human albumin.In the suspension of every milliliter of (mL) reprovision, contain 5mg paclitaxel.Effect of nano-paclitaxel injection suspension is not containing solvent, not containing floating (cremophor) (polyoxyethylene castor oil) of breast.

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: new (the romidepsin) (FK-228 of sieve meter;

), ADS-100380,

cG-781

cG-1521

sB-556629 chlamydocin

jNJ-16241199

or SAHA (vorinostat) (SAHA;

).

In one embodiment of the invention, antibody of the present invention or its Fab and etoposide (VP-16;

) combine and provide.

In one embodiment of the invention, antibody of the present invention or its Fab and gemcitabine combine and provide.

In one embodiment of the invention, disclosed any compound in antibody of the present invention or its Fab and the U.S. Patent Application No. U.S.2004/0209878A1 that announced (for example comprise by the core texture representing) or doxorubicin (

) combine and provide, comprise pattern Lay or (doxorubicin hydrochloride liposome injection; Ortho Biotech Products L.P; Raritan, NJ).

comprise

doxorubicin in liposome vectors, wherein liposome vectors is by N-(carbonyl-methoxy poly (ethylene glycol) 2000)-1, and 2-distearyl-sn-glyceryl-3-phosphoethanolamine sodium salt (MPEG-DSPE), complete all hydrogenated S-PC (HSPC) and cholesterol form.

In one embodiment of the invention, antibody of the present invention or its Fab and 5 '-deoxidation-5-fluorouracil nucleoside combine and provide.

In one embodiment of the invention, antibody of the present invention or its Fab and vincristine

combine and provide.

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: temozolomide (temozolomide)

any CDK inhibitor, ZK-304709 for example, fills in sharp Seeley (Seliciclib) (R-Loews can Wei Ting (R-roscovitine))

); Any mek inhibitor, for example PD0325901 aZD-6244; Capecitabine (capecitabine) (the fluoro-N-[(amoxy of 5 '-deoxidation-5-) carbonyl]-cytidine); Or N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo-[2,3-d] pyrimidine-5-yl) ethyl] benzoyl]-L-disodium glutamate salt heptahydrate (

pemetrexed disodium heptahydrate (pemetrexed disodium heptahydrate)).

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: camptothecine (camptothecin) (

stork etc., J.Am.Chem.Soc.93 (16): 4074-4075 (1971); Beisler etc., J.Med.Chem.14 (11): 1116-1117 (1962)); Irinotecan (

with sell; Pharmacia & Upjohn Co.; Kalamazoo, MI); Irinotecan, 5-fluorouracil (5-fluorouracil) and folinic acid (leucovorin) combination; Or the irinotecan of PEG labelling.

In one embodiment of the invention, antibody of the present invention or its Fab and FOLFOX scheme (Ao Shalipa (oxaliplatin)

together with infusion fluorouracil

and folinic acid (folinic acid)

) combine (Chaouche etc., Am.J.Clin.Oncol.23 (3): 288-289 (2000) are provided; De Gramont etc., J.Clin.Oncol.18 (16): 2938-2947 (2000)).

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with for example following estrogen antagonist: (tamoxifen (tamoxifen); By AstraZeneca Pharmaceuticals LP Wilmington, DE with

sell; ) or

(Toremifene Citrate (toremifene citrate); By Shire US, Inc.; Florence, KY with

sell).

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with for example following aromatase inhibitor:

(Anastrozole (anastrazole); By AstraZeneca Pharmaceuticals LP; Wilmington, DE with

sell), (exemestane (exemestane); By Pharmacia Corporation; Kalamazoo, MI with

sell) or

(letrozole (letrozole); By Novartis Pharmaceuticals Corporation; East Hanover, NJ with sell).

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with for example following estrogen: DES (diethylstilbestrol (diethylstilbestrol)),

(estradiol (estradiol); By Warner Chilcott, Inc.; Rockaway, NJ with sell) or conjugated estrogen hormone (conjugated estrogen) (by Wyeth Pharmaceuticals Inc.; Philadelphia, PA with

sell).

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: anti-angiogenic drugs, comprises bevacizumab (bevacizumab) (Avastin ^tM; Genentech; San Francisco, CA); Anti-VEGFR-2 antibody IMC-1C11; Other VEGFR inhibitor, for example CHIR-258

disclosed any inhibitor: WO2004/13145 in following international publication number (for example comprise core texture formula:

), WO2004/09542 (for example comprise core texture formula: ), WO00/71129 (for example comprise core texture formula:

), WO2004/09601 (for example comprise core texture formula:

), WO2004/01059 (for example comprise core texture formula:

) WO01/29025 (for example comprise core texture formula:

), WO02/32861 (for example comprise core texture formula:

) or WO03/88900 (for example comprise core texture formula ); 3-[5-(methyl sulphonyl piperidine methyl)-indyl]-quinolinones; Vatalanib (Vatalanib) (

pTK/ZK, CPG-79787, ZK-222584); AG-013736

and VEGF trap (AVE-0005), be a kind of solubility trapping receptor that comprises vegf receptor 1 and vegf receptor 2 parts.

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: LHRH (luteinising hormone-releasing hormo (Lutenizinghormone-releasing hormone)) agonist, [D-Ser (But) 6, Azgly10] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser (But)-Leu-Arg-Pro-Azgly-NH for example ₂acetate; Acetic acid [C ₅₉h ₈₄n ₁₈o ₁₄(C ₂h ₄o ₂) _x, x=1-2.4 wherein];

(goserelin acetate (goserelinacetate); By AstraZeneca UK Limited, Macclesfield, England with

sell);

(acetic acid leuproside (leuprolide acetate); By Sanofi-Synthelabo Inc.; New York, NY with

sell) or (triptorelin pamoate (triptorelin pamoate); By Pharmacia Company, Kalamazoo, MI with

sell).

In one embodiment of the invention, antibody of the present invention or its Fab and Sutent or Sunitinib malate

combine and provide.

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with for example following progestational agents (progestational agent):

(medroxyprogesterone acetate (medroxyprogesteroneacetate); By Pharmacia & Upjohn Co.; Kalamazoo, MI with

sell),

(hydroxyprogesterone caproate (hydroxyprogesterone caproate), pregnant-4-alkene-3 of 17-((1-oxo-hexyl) oxygen base), 20-diketone), megestrol acetate (megestrol acetate) or progestational hormone medicine.

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with for example following selective estrogen receptor modulators (SERM):

(raloxifene (raloxifene); By Eli Lilly andCompany; Indianapolis, IN with

sell).

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with following antiandrogen, include but not limited to:

(bicalutamide (bicalutamide); By AstraZeneca Pharmaceuticals LP; Wilmington, DE with

sell); (flutamide (flutamide); 2-methyl-N-[4-nitro-3 (trifluoromethyl) phenyl] propionic acid amide.; By Schering Corporation, Kenilworth, NJ with

sell);

(nilutamide (nilutamide); By Aventis Pharmaceuticals Inc., Kansas City, MO with

sell) and

(megestrol acetate; By Bristol-Myers Squibb with

sell).

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with the inhibitor of one or more antagonisms EGF receptor or HER2 effect, include but not limited to CP-724714

tAK-165

hKI-272

oSI-774

erlotinib, Hidalgo etc., J.Clin.Oncol.19 (13): 3267-3279 (2001)); Lapatinib (Lapatanib) (

gW2016; Rusnak etc., Molecular Cancer Therapeutics 1:85-94 (2001); The chloro-4-[(3-luorobenzyl of N-{3-) oxygen base] phenyl }-6-[5-({ [2-(methyl sulphonyl) ethyl] amino } methyl)-2-furyl]-4-quinazoline amine, PCT application number WO99/35146); Ka Na is for Buddhist nun (Canertinib) (CI-1033; erlichman etc., Cancer Res.61 (2): 739-48 (2001); Smaill etc., J.Med.Chem.43 (7): 1380-97 (2000)); ABX-EGF antibody (Abgenix, Inc.; Freemont, CA; Yang etc., Cancer Res.59 (6): 1236-43 (1999); Yang etc., Crit Rev Oncol Hematol.38 (1): 17-23 (2001)); Erbitux (erbitux) (U.S. Patent number 6,217,866; IMC-C225, Cetuximab (cetuximab); Imclone; New York, NY); EKB-569 ( wissner etc., J.Med.Chem.46 (1): 49-63 (2003)); PKI-166 (

cGP-75166); GW-572016; Any anti-egfr antibodies and any Anti-HER 2.

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing:

(chlorine Na Fani (lonafarnib); Sarasar ^tM; Schering-Plough; Kenilworth, NJ).In another embodiment, one of following fpt inhibitor is combined and is provided together with antibody of the present invention or its Fab:

Can combine with antibody of the present invention or its Fab other fpt inhibitor providing comprise BMS-214662 (

hunt etc., J.Med.Chem.43 (20): 3587-95 (2000); Dancey etc., Curr.Pharm.Des.8:2259-2267 (2002); (R)-7-cyano group-2,3,4,5-tetrahydrochysene-1-(1H-imidazol-4 yl methyl)-3-(phenyl methyl)-4-(2-thienyl sulphonyl base)-1H-1,4-benzodiazepine

)) and R155777 (Zarnestra (tipifarnib), Garner etc., Drug Metab.Dispos.30 (7): 823-30 (2002); Dancey etc., Curr.Pharm.Des.8:2259-2267 (2002); (B) amino (4-chlorphenyl) (1-methyl isophthalic acid H-imidazoles-5-yl)-methyl of-6-[]-4-(3-chlorphenyl)-1-methyl-2 (1H)-quinolinone];

with Zarnestra ^tMsell; Johnson & Johnson; New Brunswick, NJ).

In one embodiment of the invention, antibody of the present invention or its Fab and following medication combined providing: (Amifostine (Amifostine));

(NVP-LAQ824; Atadja etc., Cancer Research 64:689-695 (2004));

(suberoylanilide hyroxamic acid (suberoyl analide hydroxamic acid));

(valproic acid; Michaelis etc., Mol.Pharmacol.65:520-527 (2004));

(Trichostatin A (trichostatin A));

(FK-228; Furumai etc., Cancer Research62:49 16-4921 (2002));

(SU11248; Mendel etc., Clin.Cancer Res.9 (1): 327-37 (2003));

(BAY43-9006; Sorafenib);

(KRN951);

(aminoglutethimide (Aminoglutethimide)); (amsacrine (Amsacrine));

(anagrelide (Anagrelide));

(Anastrozole; By AstraZeneca PharmaceuticalsLP; Wilmington, DE sells with Arimidex); Asparaginase; BCG vaccine (BacillusCalmette-Guerin, BCG) vaccine (Garrido etc., Cytobios.90 (360): 47-65 (1997));

(bleomycin (Bleomycin));

(Buserelin (Buserelin));

(busulfan (Busulfan); Two methanesulfonic acid BDOs; By ESP Pharma, Inc.; Edison, New Jersey with

Sell);

(carboplatin; By Bristol-Myers Squibb; Princeton,NJ with

Sell);

(BCNU (Carmustine));

(Chlorambucil (Chlorambucil)); (cis-platinum);

(Cladribine (Cladribine));

(clodronate (Clodronate));

(endoxan);

(cyproterone (Cyproterone));

(cytarabine (Cytarabine)); (Dacarbazine (Dacarbazine));

(actinomycin D);

(daunorubicin (Daunorubicin));

(diethylstilbestrol);

(epirubicin (epirubicin)); (fludarabine (Fludarabine));

(fludrocortison (Fludrocortisone));

(Fluoxymesterone (Fluoxymesterone));

(Flutamide); (hydroxycarbamide (Hydroxyurea));

(idarubicin (Idarubicin));

(ifosfamide); (Imatinib (Imatinib); By Novartis PharmaceuticalsCorporation; East Hanover, NJ with

Disappear and sell);

(folinic acid);

(leuproside);

(levamisol (Levamisole));

(lomustine (Lomustine));

(mustargen (Mechlorethamine));

(melphalan (Melphalan); By Celgene Corporation; Warren, NJ with

Sell);

(purinethol (Mercaptopurine));

(mesna (Mesna));

(methotrexate (MTX));

(mitomycin (Mitomycin)); (mitotane (Mitotane));

(mitoxantrone (Mitoxantrone));

(Nilutamide); Octreotide (octreotide) (

Katz etc., Clin Pharm.8 (4): 255-73 (1989); Sandostatin

Durative action preparation; Novartis Pharm.Corp; E.Hanover, NJ); According to writing music peptide (edotreotide) (Yttrium-90 mark or unlabelled); Ao Shalipa (

Sanofi-Synthelabo Inc.; New York, NY is with Eloxatin ^TMBy selling); (Pamidronate (Pamidronate); By Novartis Pharmaceuticals Corporation; East Hanover, NJ with

Sell);

(Pentostatin (Pentostatin); By Supergen; Dublin, CA with

Sell);

(plicamycin (Plicamycin));

(porphines nurse (Porfimer); By Axcan Scandipharm Inc.; Birmingham, AL with

Sell);

(procarbazine (Procarbazine));

(Raltitrexed (Raltitrexed)); Rituximab (Rituximab) (by Genentech, Inc.; South San Francisco, CA with

Sell);

(streptozotocin (Streptozocin));

(Teniposide); (testosterone (Testosterone));

(Thalidomide (Thalidomide));

(thioguanine (Thioguanine));

(phosphinothioylidynetrisaziridine (Thiotepa));

(Tretinoin (Tretinoin)); (eldisine (Vindesine)) or 13-cisRA

In one embodiment of the invention, antibody of the present invention or its Fab and following any one or more medication combined providing: melphalan (phenylalaninemustard), uracil mustard (uracil mustard), estramustine (estramustine), altretamine (altretamine), floxuridine (floxuridine), 5-FU (5-deooxyuridine), cytosine arabinoside (cytosine arabinoside), Ismipur (6-mecaptopurine), deoxycoformycin (deoxycoformycin), calcitriol (calcitriol), valrubicin (valrubicin), mithramycin (mithramycin), vinblastine (vinblastine), vinorelbine (vinorelbine), hycamtin (topotecan), razoxin, Marimastat (marimastat), COL-3, Neovastat (neovastat), BMS-275291, amine (squalamine) is overstated by department, endostatin (endostatin), SU5416, SU6668, EMD121974, IL-12 (interleukin-12), IM862, angiostatin (angiostatin), the α V β anti-monoclonal antibody of 3 humanization (vitaxin), droloxifene (droloxifene), indoxifene (idoxyfene), spironolactone (spironolactone), finasteride (finasteride), cimetidine (cimitidine), Herceptin (trastuzumab), denileukin (denileukin), diphtheria toxin, diphtherotoxin transmembrane segment (diftitox), gefitinib (gefitinib), bortezomib (bortezimib), paclitaxel, docetaxel (docetaxel), epothilone B (epithilone B), BMS-247550 (referring to such as Lee etc., Clin.Cancer Res.7:1429-1437 (2001)), BMS-310705, droloxifene (3-trans-Hydroxytamoxifen (3-hydroxytamoxifen)), 4-hydroxytamoxifen, ERA 923 (pipendoxifene), ERA-923, arzoxifene (arzoxifene), fulvestrant (fulvestrant), acolbifene (acolbifene), lasofoxifene (lasofoxifene) (CP-336156), idoxifene (idoxifene), TSE-424, HMR-3339, ZK186619, hycamtin, PTK787/ZK 222584 (Thomas etc., Semin Oncol.30 (the 3rd phase supplementary issue 6): 32-8 (2003)), humanization VEGF antibody bevacizumab, VX-745 (Haddad, Curr Opin.Investig.Drugs 2 (8): 1070-6 (2001)), PD 184352 (Sebolt-Leopold etc., Nature Med.5:810-816 (1999)), any mTOR inhibitors, rapamycin (rapamycin) (

sirolimus (sirolimus)), 40-O-(2-ethoxy)-rapamycin, CCI-779 (

cCI-779 (temsirolimus), Sehgal etc., Med.Res.Rev., 14:1-22 (1994), 1249-53 (2002)), AP-23573 Elit, Curr.Opin.Investig.Drugs 3 (8):

rAD001

aBT-578

bC-210 lY294002, LY292223, LY292696, LY293684, LY293646 (Vlahos etc., J.Biol.Chem.269 (7): 5241-5248 (1994)), wortmannin (wortmannin), BAY-43-9006 (Wilhelm etc., Curr. Pharm.Des.8:2255-2257 (2002)), ZM336372, L-779,450, (L86-8275/HMR 1275 for the disclosed any Raf inhibitor of people (Curr.Pharm Des.8:2269-2278 (2002)), the flavone pyrrole many (flavopiridol) such as Lowinger, Senderowicz, Oncogene 19 (56): 6600-6606 (2000))

In one embodiment of the invention, antibody of the present invention or its Fab are combined and are provided with any one or more compound described in following patent: United States Patent (USP) 5,656,655, and it discloses the heteroaryl EGFR inhibitor that styryl replaces; United States Patent (USP) 5,646,153, it discloses two monocycles and/or aryl bicyclic heteroaryl carbocyclic ring and assorted carbocyclic ring EGFR inhibitor and PDGFR inhibitor; United States Patent (USP) 5,679,683, it discloses the tricyclic pyrimidine compound that suppresses EGFR; United States Patent (USP) 5,616,582, it discloses has receptor tyrosine kinase and suppresses active quinazoline derivant; Fry etc., Science 2651093-1095 (1994), it discloses the compound (referring to Fig. 1 of the people such as Fry) with the structure that suppresses EGFR; United States Patent (USP) 5,196,446, it discloses heteroaryl ethylene two bases or the heteroaryl ethylene two base aryl compounds that suppress EGFR; Panek etc., Journal of Pharmacology and ExperimentalTherapeutics 283:1433-1444 (1997), it disclose compound be accredited as suppress Receptor EGFR, PDGFR and FGFR family PD166285---PD166285 is accredited as 6-(2,6-Dichlorobenzene base)-2-(4-(2-diethylamino ethoxy) phenyl amino)-8-methyl-8H-pyrido (2,3-d) pyrimidin-7-ones.

In one embodiment of the invention, antibody of the present invention or its Fab and following any one or more medication combined providing: Pegylation or not glycol interferon alpha-2a, Pegylation or not glycol interferon alpha-2b, Pegylation or not glycol interferon alpha-2c, Pegylation or not glycol interferon alpha n-1, Pegylation or not glycol interferon alpha n-3 and Pegylation, not Pegylation interferon alfacon-1 or albumin interferon-' alpha '.

Term used herein " interferon-ALPHA " refers to the species specificity protein family that suppresses cell proliferation and regulate the height homology of immunne response.Suitable interferon-' alpha ' generally includes but is not limited to Interferon Alfa-2b, Interferon Alfa-2a, recombinantinterferonα-2c, α 2 interferon, interferon alfa-n1 (INS), pure natural interferon alpha mixture, interferon alfacon-1 (referring to for example U.S. Patent number 4,897,471 and 4,695,623 (embodiment 7,8 or 9 especially wherein)), the mixture of Alferon N or natural interferon alpha.

Intederon Alpha-2a by Hoffmann-La Roche (Nutley, N.J.) with

sell.

Interferon Alpha-2b by Schering Corporation (Kenilworth, NJ) with

sell.The preparation of interferon alpha 2 b is referring to for example U.S. Patent number 4,530,901.

Alferon N is the mixture of natural interferon, by Hemispherx Biopharma, Inc. (Philadelphia, PA) with sell.

Interferon alfa-n1 (INS) is the mixture of natural interferon, by Glaxo-Smith-Kline (Research Triangle Park, NC) with

sell.

Interferon alfacon-1 is by Intermune, Inc. (Brisbane, CA) with sell.

Interferon α-2 c is by Boehringer Ingelheim Pharmaceutical, Inc. (Ridgefield, CT) with

sell.

The mixture of pure natural interferon is by Sumitomo; Tokyo, Japan with sell.

Term used herein " glycol interferon alpha " refers to polyethyleneglycol modified interferon alpha conjugate, preferably Intederon Alpha-2a and Interferon Alpha-2b.Preferred Polyethylene Glycol Interferon Alpha-2b conjugate is PEG 12000-Interferon Alpha-2b.Term used herein " 12,000 molecular weight polyisoprene ethylene glycol conjugated interferon α " and " PEG 12000-IFN α " comprise for example according to containing the conjugate that urethane bonds and Polyethylene Glycol mean molecule quantity are 12000 between the method preparation of international application no WO95/13090 and EP1039922 and Intederon Alpha-2a or Interferon Alpha-2b amino.Glycol interferon alpha, PEG 12000-IFN-α-2b can be available from Schering-Plough Research Institute, Kenilworth, N.J..

Preferred PEG 12000-Interferon Alpha-2b can be by being connected PEG polymer to prepare with the histidine residues on Interferon Alpha-2b molecule.Single PEG 12000 molecules can be puted together by the free amine group on urethane bonds and IFN α-2b molecule.The feature of this conjugate is that to be connected with molecular weight be 12000 PEG.PEG 12000-IFN α-2b conjugate can be made to the lyophilized injectable powder for injecting.

Glycol interferon alpha-2b by Schering Corporation (Kenilworth, NJ) with

sell.

Glycol interferon-α-2a by Hoffmann-La Roche (Nutley, N.J.) with sell.

Other interferon alpha conjugate can be by preparing interferon-ALPHA and water-soluble polymer coupling.The non-limitative example of this base polymer comprises other polyalkylene oxide homopolymer, for example polypropylene glycol, polyoxyethylene polyhydric alcohol, its copolymer and its block copolymer.As the alternative compounds of the polymer based on polyalkylene oxide, can use effective nonantigenic material, such as glucosan, polyvinylpyrrolidone, polyacrylamide, polyvinyl alcohol, polymer based on sugared etc.This interferoid α-polymer conjugate can be referring to U.S. Patent number 4,766,106 for example, U.S. Patent number 4,917,888, European Patent Application No. 0 236 987 or 0 593 868 or international publication number WO 95/13090.Preferred PEG12000-IFN α 2b can be by being connected PEG polymer to prepare with the histidine residues on Interferon Alpha-2b molecule.

The pharmaceutical composition that is suitable for the glycol interferon alpha of parenteral can for example, for example, for example, for example, for example, for example, for example, be prepared together with suitable buffer (Tris-HCl, acetate or phosphate, dibastic sodium phosphate/phosphate sodium dihydrogen buffer solution), pharmaceutically acceptable excipient (sucrose), carrier (albumin human), toxic agents (NaCl), antiseptic (thimerosal (thimerosol), cresol or benzyl alcohol) and the surfactant (tween (tween) or Polysorbate) that is dissolved in Injectable sterile water.Glycol interferon alpha can lyophilized injectable powder stored refrigerated at 2 ℃-8 ℃.It is stable that reprovision aqueous solution is used when being kept between 2 ℃ and 8 ℃ and in 24 hours of reprovision.Referring to for example U.S. Patent number 4,492,537,5,762,923 and 5,766,582.Reprovision aqueous solution can also be at the syringe of pre-fill multiple dose, such as preserving for the syringe of the medicines such as insulin delivery.Suitable syringe generally includes such system, and this system comprises the pre-fill bottle being connected with pen-type injector, for example can be available from Novo Nordisk's

novo Pen or can be available from Schering Corporation, Kenilworth, NJ's other injector system comprises the pen-type injector that glass syringe is housed, and at syringe, independently in compartment, diluent and lyophilizing glycol interferon alpha powder is housed.

Scope of the present invention also comprises treatment or prevents the method for medical conditions described herein or disease, the method for example, by comprising the compositions of anti-IGF1R antibody of the present invention or its Fab and one or more other anticancer chemotherapeutic agent (anticancer chemotherapeutic agent described herein) and/or one or more Bendectins, Bendectin includes but not limited to Carcel smooth (casopitant) (GlaxoSmithKline), Netupitant (Netupitant) (MGI-Helsinn) and other nk 1 receptor antagonist, palonosetron (palonosetron) (being sold with Aloxi by MGI Pharma), aprepitant (aprepitant) is (by Merck and Co., Rahway, NJ sells with Emend), diphenhydramine (diphenhydramine) is (by Pfizer, New York, NY with

sale), hydroxyzine (hydroxyzine) is (by Pfizer, New York, NY with

sell), metoclopramide (metoclopramide) (by AH Robins Co, Richmond, VA with

sale), lorazepam (lorazepam) is (by Wyeth, Madison, NJ with

sale), alprazolam (alprazolam) is (by Pfizer, New York, NY with

sale), haloperidol (haloperidol) is (by Ortho-McNeil, Raritan, NJ with

sale), droperidol (droperidol)

dronabinol (dronabinol) is (by Solvay Pharmaceuticals, Inc., Marietta, GA with

sale), dexamethasone (dexamethasone) is (by Merck and Co., Rahway, NJ with

sale), methylprednisolone (methylprednisolone) is (by Pfizer, New York, NY with

sale), prochlorperazine (prochlorperazine) is (by Glaxosmithkline, Research Triangle Park, NC with

sale), granisetron (granisetron) is (by Hoffmann-La Roche Inc., Nutley, NJ with

sale), ondansetron (ondansetron) is (by Glaxosmithkline, Research Triangle Park, NC with

sale), dolasetron (dolasetron) is (by Sanofi-Aventis, New York, NY with sale), tropisetron (tropisetron) is (by Novartis, EastHanover, NJ with

sell).

The compositions that comprises Bendectin can be used for prevention or treatment is felt sick; This is the common side effect of anti-cancer chemotherapy.Therefore, the present invention includes the method that is used for the treatment of or prevents curee's cancer, the method for example, by giving antibody of the present invention or its Fabs optional and (as herein described) coupling of one or more other chemotherapeutics and/or optional and one or more Bendectin couplings.

Other side effect for the treatment of of cancer comprises erythrocyte and aleucemia.Therefore, the present invention includes and comprise anti-IGF1R antibody or the optional compositions with treating or preventing the drug combination of this class deficiency disease of its Fab, for example, such as filgrastim (pegfilgrastim), erythropoietin (erythropoietin), Epoetin Alfa (epoetin alfa) or darbepoetin α (darbepoetin alfa).

Another side effect of anti-cancer therapies is mucositis.Therefore, the invention provides the method and composition that is used for the treatment of or prevents medical conditions described herein or disease, the method gives anti-IGF1R antibody or its Fab and anti-mucositis and treats by combining.For example, catarrhal treatment comprises rinsing the mouth anesthesia viscogel (anesthetic gel viscous)

(lignocaine (lidocaine)) 2% or collutory, collutory comprises allopurinol (allopurinol), benzydamine hydrochloride (benzydamine hydrochloride), cortical steroid (corticosteroids) and/or Chamomile (chamomile).

The present invention also comprises and is used for the treatment of or prevents any stadium of any medical conditions described herein or the method for type, and the method is by giving antibody of the present invention or its Fab and treatment perform the operation for example surgery tumorectomy or anticancer radiotherapy coupling; Optionally with other chemotherapeutics as above and/or Bendectin coupling.

Term " with ... coupling or with ... associating " refer to and each component of the present composition (for example anti-IGF1R antibody or its Fab are together with docetaxel) can be mixed with to single compositions for pass medicine simultaneously, or be mixed with respectively two or more compositionss (for example medicine box).In addition, can within the time different from giving other component, give curee each component; For example, each administration can be within preset time for example, gives by the non-while of some intervals (separate or sequential).For example, and independent component can give by identical or different approach curee (wherein anti-IGF1R antibody gives through parenteral, and gefitinib oral administration gives).

Therapeutic Method and administration

The present invention includes the method that is used for the treatment of or prevents following any disease of using the optional and other chemotherapeutics of IGF1R inhibitor or its pharmaceutical dosage form coupling: incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma, chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, Burkitt lymphoma, mycosis fungoides, seary's syndrome, cutaneous T cell lymphoma, chronic myeloproliferative diseases) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor, hepatocarcinoma, gastric cancer, gigantism, pituitary adenoma, psoriasis and kidney rhabdoid tumor.

The pharmaceutical composition that comprises IGF1R inhibitor and other chemotherapeutics (for example as mentioned above) coupling and pharmaceutically acceptable carrier all falls within the scope of the present invention (for example single compositions or be contained in medicine box respectively).Pharmaceutical composition can be prepared by the well-known any method of pharmaceutical field; Referring to such as (chief editor) (1990) such as Gilman, the PharmacologicalBases of Therapeutics, the 8th edition, Pergamon Press; A.Gennaro (chief editor), remington ' s Pharmaceutical Sciences, the 18th edition, (1990), Mack PublishingCo., Easton, Pennsylvania.; Avis etc. (chief editor) (1993) pharmaceutical DosageForms:Parenteral Medicationsdekker, New York; Lieberman etc. (chief editor) (1990) pharmaceutical Dosage Forms:Tabletsdekker, New York; With (chief editor) (1990) such as Lieberman, pharmaceutical Dosage Forms:DisperseSystemsdekker, New York.In one embodiment of the invention, antibody gives curee as the ingredient of pharmaceutical composition, this pharmaceutical composition comprises antibody, for example LCF/HCA (for example 1mg/ml, 5mg/ml, 10mg/ml, 15mg/ml, 20mg/ml or 25mg/ml); Sodium acetate trihydrate (for example 1.8mg/ml, 2.3mg/ml or 3.1mg/ml); Glacial acetic acid (for example 0.5mg/ml, 0.18mg/ml or 2.2mg/ml); Sucrose (for example 50mg/ml or 70mg/ml) and water (pH approximately 5.5).

Term " incidence cancer " etc. comprises any cancer that occurs in health head and neck position, comprises the cancer of oral cavity, pharynx, nasal sinuses, nasal cavity, larynx and salivary gland (for example gland, sublingual gland under the parotid gland, lower volume).A general type of incidence cancer is the squamous cell carcinoma that occurs in head and neck position.Other type of incidence cancer comprises salivary tumor transformation, melanoma, lymphoma and sarcoma.

Term " squamous cell carcinoma " (SCC) etc. comprises and involves the squamocellular any cancer of skin.This term comprises any hypotype of squamous cell carcinoma, for example comprises keratoacanthoma, wears pick property cancer (carcinoma cuniculatum), aggressive SCC, original position SCC or transitivity SCC.

Term " multiple myeloma " and " solitary plasmacytoma " etc. comprise plasma cell cancer.Solitary plasmacytoma is the myeloma at single position in body.Term " multiple myeloma " for example comprises the wherein generation myeloma cell's at a plurality of positions (for example bone marrow position) embodiment in vivo.This term comprises any hypotype of multiple myeloma, comprises for example light chain type myeloma, nonsecreting type myeloma.

Term " renal cell carcinoma " etc. is also called kidney portion cancer or renal adenocarcinoma, comprises the embodiment of the cancerous cell in any tissue that is wherein present in kidney.This term comprises for example any hypotype of renal cell carcinoma, comprises for example clear cell carcinoma (no matter whether mixing with granulocyte), chromatophile cancer (chromophobic cancer), kidney Rhabdoid tumor, chromophobe cell tumor, acidophil carcinoma, Collecting duct carcinoma, transitional cell carcinoma and sarcoma sample tumor.

Term " curee " or " patient " comprise any biology, and preferred mammal (for example primates, Canis familiaris L., horse, rat, mice, cat, rabbit), is most preferably people.In one embodiment of the invention, " curee " or " patient " for adult's (for example 18 years old or more than) or child (under-18s for example, for example 1 years old following, 1 years old, 2 years old, 3 years old, 4 years old, 5 years old, 6 years old, 7 years old, 8 years old, 9 years old or 10 years old).

The pharmaceutical composition that can use conventional pharmaceutically acceptable excipient and additive and routine techniques preparation to contain the optional and other chemotherapeutics coupling of IGF1R inhibitor.The pharmaceutically acceptable excipient of this class and additive comprise nontoxic compatible filler, binding agent, disintegrating agent, buffer agent, antiseptic, antioxidant, lubricant, correctives, thickening agent, coloring agent, emulsifying agent etc.The present invention includes all route of administration, include but not limited to parenteral (for example subcutaneous, intravenous, intraperitoneal, intramuscular, part, intraperitoneal, suction, intracranial) and non-parenteral (for example oral, percutaneous, intranasal, ophthalmic, Sublingual, rectum and part).

Injection can conventionally form preparation, as liquid solution agent or suspensoid, be suitable for being dissolved in before injection the solution of liquid or the solid form of suspensoid or as Emulsion.Injection, solution and Emulsion also can contain one or more excipient.Excipient comprises for example water, saline, glucose, glycerol or ethanol.In addition, if needed, pharmaceutical composition to be given also can contain a small amount of nontoxic auxiliary substance, for example wetting agent or emulsifying agent, pH buffer agent, stabilizing agent, solubilizing agent and other composition, for example sodium acetate, sorbitan laurate, triethanolamine oleate and cyclodextrin.

In one embodiment of the invention, the pharmaceutically acceptable carrier for parenteral formulation comprises aqueous vehicle, non-water-soluble matchmaker, antimicrobial, isotonic agent, buffer agent, antioxidant, local anesthetic, suspension dispersive agent, emulsifying agent, sequestering agent or chelating agen and other pharmaceutically acceptable material.

The example of aqueous vehicle comprise sodium chloride injection, ringer's inj, etc. ooze glucose injection, sterilized water injection, glucose and lactate ringer's inj.Non-water parenteral solvent comprises fixedly oil, Oleum Gossypii semen, Semen Maydis oil, Oleum sesami and the Oleum Arachidis hypogaeae semen of plant source.Must, to the antimicrobial that adds anti-bacteria concentration or Antifungi concentration in the parenteral formulation being packaged in multi-dose container, comprise phenol or cresol, mercurial, benzyl alcohol, chlorobutanol, methyl parahydroxybenzoate, propyl p-hydroxybenzoate, thimerosal, benzalkonium chloride and benzethonium chloride.Isotonic agent comprises sodium chloride and glucose.Buffer agent comprises phosphate and citrate.Antioxidant comprises sodium bisulfate.Local anesthetic comprises procaine hydrochloride.Suspension dispersive agent comprises sodium carboxymethyl cellulose, hydroxypropyl emthylcellulose and polyvinylpyrrolidone.Emulsifying agent comprises polyoxyethylene sorbitan monoleate (TWEEN-80).Sequestering agent or the chelating agen of metal ion comprise EDTA.Pharmaceutical carrier also comprises for the ethanol of water miscibility solvent, Polyethylene Glycol and propylene glycol; And for sodium hydroxide, hydrochloric acid, citric acid or the lactic acid of pH regulator.

In one embodiment of the invention, for the preparation of parenteral can comprise ready-made for the sterile solution agent of injecting, before use easily with the aseptic anhydrous soluble products (for example lyophilized injectable powder) of solvent, comprise hypodermic tablet, ready-made for the aseptic suspensoid injected, easy mix with solvent aseptic without water-insoluble product and aseptic Emulsion before use.Solution can be aqueous pharmaceutical or non-aqueous solution agent.

Also comprise implantation slow release (slow-release/sustained-release) system herein so that dosage level keeps constant.Say simply, activating agent (IGF1R inhibitor for example, optional and other chemotherapeutics coupling) is scattered in solid interior substrate, internal matrix is insoluble in the outside polymeric membrane of body fluid and surrounds, and solid interior substrate is polymethyl methacrylate for example, polybutyl methacrylate, plasticizing or not plastized polyvinyl chloride, plasticizing nylon, plasticizing polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, vinyl-vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, siloxanes carbonic ester copolymer, hydrophilic polymer (for example hydrogel of acrylate and methacrylate), collagen, cross-linking polyvinyl alcohol and crosslink part hydrolyzed poly vinyl acetate, outside polymeric membrane is polyethylene for example, polypropylene, ethylene/propene copolymer, ethylene/ethyl acrylate copolymer, ethylene/vinyl acetate, silicone rubber, polydimethylsiloxane, neoprene, chlorinated polyethylene, polrvinyl chloride, the copolymer of vinyl chloride and vinyl acetate/vinylidene chloride/ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene/vinyl alcohol copolymer, Ethylene/vinyl acetate/vinyl alcohol terpolymer and ethylene/vinyl ethoxy-ethanol copolymer.In rate of release, control in step, compound spreads apart by outer polymeric membrane.The percentage ratio of the reactive compound comprising in this parenteral compositions depends on its special properties to a great extent, and the activity of IGF1R inhibitor and curee's the needs of optional and other chemotherapeutics coupling.

Can regulate the concentration of the IGF1R inhibitor of optional and other chemotherapeutics coupling, make a shot that the effective dose that produces required pharmacological action can be provided.As discussion below, definite dosage depends on age, body weight and the general status of patient or animal, as known in the art.

In one embodiment of the invention, unit dose parenteral formulation is contained in ampoule, bottle or band needle injection.All preparations for parenteral must be aseptic, as known in the art and enforcement.

In one embodiment of the invention, by the optional and other chemotherapeutics coupling of IGF1R inhibitor and be mixed with lyophilized injectable powder, this lyophilized injectable powder can be usingd as solution, Emulsion and other mixture administration by reprovision.Injectable powder also can be made solid or gel by reprovision.

In one embodiment of the invention, for example, by being dissolved in, the IGF1R inhibitor (anti-IGF1R antibody) of optional and other chemotherapeutics coupling or its pharmaceutically acceptable derivates in suitable solvent, prepare aseptic freeze-dried injectable powder.This solvent can be containing the reprovision solution that is improved the excipient of stability or the other medicines component of injectable powder or is prepared by injectable powder.Operable excipient includes but not limited to glucose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose or other suitable composition.This solvent also can contain buffer agent, and for example citrate, sodium phosphate or potassium phosphate or other this class buffer agent known in the art, in one embodiment, be about neutral pH.Subsequently under the known standard conditions of those skilled in the art, make lyophilizing after this solution filtration sterilization, obtain required preparation.In one embodiment, gained solution is divided and installed in bottle with lyophilizing.Each bottle can be equipped with the IGF1R inhibitor of the optional and other chemotherapeutics coupling of single dose or multiple dose.It is acceptable that bottle overfilling surpasses on a small quantity required dosage or dosage group (for example approximately 10%), makes to be conducive to accurate draw samples and accurately administration.Can for example, at the lower lyophilized injectable powder of preserving of suitable condition (approximately 4 ℃ to room temperature).

Lyophilized injectable powder and water for injection reprovision obtain the dosage form for parenteral.In one embodiment of the invention, during reprovision, lyophilized injectable powder is added in sterilized water or other suitable carrier.Accurate dosage depends on the selected therapy to be supplied that has.This consumption can be determined by rule of thumb.

Can use the aerosol of Sorbitan Trioleate or oleic acid and the dose regimen providing together with for example Arcton 11, dichlorofluoromethane, dichlorotetra-fluoroethane or any other biocompatibility impelling gas by sucking are for example provided; Also likely utilize the system of injectable powder form, this system contain the optional and other chemotherapeutics coupling of IGF1R inhibitor itself or with excipient coupling.

In one embodiment of the invention, the chemotherapeutics coupling that IGF1R inhibitor is optional and other, is made into the solid dosage forms for oral administration, in one embodiment, makes capsule or tablet.Tablet, pill, capsule, buccal tablet (troch) etc. can contain one or more following ingredients or have the compound of similarity: binding agent, lubricant, diluent, fluidizer, disintegrating agent, coloring agent, sweeting agent, correctives, wetting agent, enteric coating (emetic coating) and film coating.The example of binding agent comprises microcrystalline Cellulose, tragakanta, glucose solution, mucialga of arabic gummy agent, gelatin solution, molasses, polyvinylpyrrolidine, polyvidone, polyvinylpolypyrrolidone, sucrose and starch slurry.Lubricant comprises Pulvis Talci, starch, magnesium stearate, calcium stearate, lycopodium and stearic acid.Diluent comprises for example lactose, sucrose, starch, Kaolin, salt, mannitol and dicalcium phosphate.Fluidizer includes but not limited to colloidal silica.Disintegrating agent comprises that cross-linked carboxymethyl cellulose is received, sodium starch glycollate, alginic acid, corn starch, potato starch, Bentonite, methylcellulose, agar and carboxymethyl cellulose.Coloring agent comprises water solublity FD and C dyestuff, its mixture that for example any empirical tests is got permission and is suspended in water-insoluble FD and the C dyestuff in alumina hydrate.Sweeting agent comprises sucrose, lactose, mannitol and artificial sweetening agent (for example dry spice of glucide and multiple spraying).Correctives comprises the natural perfume material for example, extracting from plant (fruit) and the mixture that produces the synthetic compound of comfort, such as but not limited to Oleum menthae and methyl salicylate.Wetting agent comprises propylene glycol monostearate, sorbitan monooleate, diglycol laurate and polyoxyethylene dodecylic acid ether.Enteric coating comprises fatty acid, fat, wax, Lac, ammonification Lac and cellulose acetate phthalate.Film coating comprises hydroxyethyl-cellulose, sodium carboxymethyl cellulose, Macrogol 4000 and cellulose acetate phthalate.

Any composition as herein described can be mixed with slow release formulation and comprise Lipidosome, for example unilamellar liposome (ULV), multilamellar liposome (MLV) and DepoFoam ^tMgranule (Kim etc., Biochim.Biophys.Acta (1983) 728 (3): 339-348; Kim, Methods Neurosci. (1994) 21:118-131; Kim etc., Anesthesiology (1996) 85 (2): 31-338; Katre etc., J.Pharm.Sci. (1998) 87 (11): 1341-1346).The feature of DepoFoam system is, has the discontinuous inside of being separated by the liposome membrane of non-concentric continuous net containing hydroecium in each DepoFoam granule, provide higher moisture volume/lipid than and compare much bigger particle diameter with MLV.

Dosage and administration

Method of the present invention comprises IGF1R inhibitor or its pharmaceutical composition that gives optional and other chemotherapeutics coupling.If possible, conventionally just according to physicians ' DeskReference 2003( physicians ' Desk Reference, the 57th edition); MedicalEconomics Company; ISBN:1563634457; In the 57th edition (in November, 2002), check and approve scheme listed in the product description of medicine and therapeutic scheme well-known in the art, implement administration and the dosage of this class medicine.

Term " treatment effective dose " or " treatment effective dose " refer to (research worker for example by implementer, doctor or veterinary) definite can induced tissue, system, amount or the dosage of curee or host's biological respinse or the present composition of medical response (for example IGF1R inhibitor, for example anti-IGF1R antibody), described reaction comprises any measurable for example sign of following cancer that alleviates in any degree, symptom and/or clinical marker (comprise and for example suppress any IGF 1R activity, for example IGF-I or IGF-II are in conjunction with activity or kinase activity) and/or prevention, slow down or stop process or the transfer of described cancer: incidence cancer, squamous cell carcinoma, multiple myeloma, solitary plasmacytoma, renal cell carcinoma, retinoblastoma, germ cell tumor, hepatoblastoma, hepatocarcinoma, melanoma, kidney rhabdoid tumor, Ewing sarcoma, chondrosarcoma, any hematologic malignancies (chronic lymphoblast leukemia for example, leukemia chronic myelo-monocytic, acute lymphoblast leukemia (T cell lineage for example, B cell precursor pedigree), acute lymphoblastic leukemia, acute myelocytic leukemia, acute myeloblast leukemia, chronic myeloblast leukemia, Hodgkin, non-Hodgkin lymphoma, chronic lymphocytic leukemia, chronic myelocytic leukemia, myelodysplastic syndrome, hairy cell leukemia, mast cell leukemia, mastocytoma, follicular lymphoma, diffuse large cell lymphoma, lymphoma mantle cell, Burkitt lymphoma, mycosis fungoides, seary's syndrome, cutaneous T cell lymphoma, chronic myeloproliferative diseases) and the central nerve neuroma (brain cancer for example, glioblastoma, the non-glioblastoma brain cancer, meningioma, pituitary adenoma, vestibular schwannomas, intramedullary primitive neuroectodermal tumor, medulloblastoma, astrocytoma, glioblastoma multiforme, oligodendroglioma, ependymoma and papilloma of choroid plexus), myeloproliferative diseases (for example polycythemia vera, thrombocytosis, idiopathic myelofibrosis), soft tissue sarcoma, thyroid carcinoma, carcinoma of endometrium, carcinoid, germ cell tumor and hepatocarcinoma (for example tumor growth or cancer cell multiplication).For example, in one embodiment, " the treatment effective dose " of any anti-IGF1R antibody, for example, comprise the antibody of following variable region or " the treatment effective dose " of its Fab or any other anti-IGF1R antibody as herein described is twice weekly, once in a week, once every two weeks, every three weeks once or January once, each consumption is (about 0.5mg/kg body weight for example between about 0.3mg/kg body weight and about 20mg/kg body weight, about 1mg/kg body weight, about 2mg/kg body weight, about 3mg/kg body weight, about 4mg/kg body weight, about 5mg/kg body weight, about 6mg/kg body weight, about 7mg/kg body weight, about 8mg/kg body weight, about 9mg/kg body weight, about 10mg/kg body weight, about 11mg/kg body weight, about 12mg/kg body weight, about 13mg/kg body weight, about 14mg/kg body weight, about 15mg/kg body weight, about 16mg/kg body weight, about 17mg/kg body weight, about 18mg/kg body weight, about 19mg/kg body weight, about 20mg/kg body weight): the variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that (a) comprises SEQ ID NO:2 and the aminoacid 20-137 that comprises SEQ ID NO:10 or 12, (b) variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that comprises SEQ ID NO:4 and the aminoacid 20-137 that comprises SEQID NO:10 or 12, (c) variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that comprises SEQ ID NO:6 and the aminoacid 20-137 that comprises SEQ ID NO:10 or 12, or the variable region of heavy chain of the variable region of light chain of the aminoacid 20-128 that (d) comprises SEQ ID NO:8 and the aminoacid 20-137 that comprises SEQ ID NO:10 or 12.

For example can adjust dosage, so that optimal reaction (therapeutic response) to be provided.For example, can give single dose, or can within a period of time, give some divided doses, or can suitably reduce or increase in accordance with regulations dosage according to emergency treatment situation.For example, ordinary skill practitioner (for example doctor or veterinary) can be according to patient age, body weight, height, passing medical history, now by medicine and potential cross reaction, allergy, sensitivity and toxic and side effects, determine or adjust dosage.Especially be advantageously mixed with the unit dosage forms of parenteral compositions, to be easy to administration and dosage homogeneity.

The practitioner (for example doctor or veterinary) with ordinary skill can easily determine the required pharmaceutical composition of effective dose and output prescription.For example, doctor or veterinary can, by starting to give for the antibody of the present invention of pharmaceutical composition or the dosage of Fab lower than reaching the needed level of ideal treatment, increase dosage until reach required effect gradually.Can whether dwindle or stop growing by for example measuring tumor to be treated in curee's body, determine the curative effect of given dose or the therapeutic scheme of antibody of the present invention or combination.Can easily measure the size of tumor, for example, by estimating or use hands palpation in X ray, nuclear magnetic resonance (MRI), operation process.Tumor size and propagation also can be applied thymidine PET scanning (thymidine PETscan) and measure (referring to such as Wells etc., Clin.Oncol.8:7-14 (1996)).Thymidine PET scanning generally comprises injection radiotracer ([2-for example ¹¹c]-thymidine), then patient body is carried out to PET scanning (Vander Borght etc., Gastroenterology 101:794-799,1991; Vander Borght etc., J.Radiat.Appl.Instrum. A part, 42:103-104 (1991)).Operable other tracer comprise [ ¹⁸f]-FDG (18-fluorodeoxyglucose), [ ¹²⁴i] IUdR (5-[124I] iodo-2 '-BrdU), [ ⁷⁶br] BrdUrd (bromodeoxyribouridine), [ ¹⁸f] FLT (3 '-deoxidation-3 ' fluorothymidine) or [ ¹¹c] FMAU (2 '-fluoro-5-methyl isophthalic acid-β-D-furan ara-U).

For example, can pass through the process of monitored in various ways incidence cancer by doctor or veterinary, and can change thus dosage.The method of monitor head cervical region cancer comprises physical examination for example (for example visual inspection or touch check), is difficult to endoscopy (for example electric nasopharyngoscope inspection, pharyngoscopy or laryngoscopy), computerization X-ray tomoscan (CT), NMR (Nuclear Magnetic Resonance)-imaging scanning (MRI), ultrasonic examination, positron emission tomography (PET) scanning, the panorex (jaw X ray) of look-out station, gulps down barium inspection, tooth X ray, chest X-ray and radionuclide bone scan.

For example, can pass through the process of monitored in various ways squamous cell carcinoma by doctor or veterinary, and can change thus dosage.The method of monitoring squamous cell carcinoma comprises for example for example, by patient's inquiring or physical examination (data logging of visual inspection and any pathological changes size, shape or other presentation quality).

For example, can pass through the process of monitored in various ways multiple myeloma by doctor or veterinary, and can change thus dosage.The method of monitoring multiple myeloma comprises for example to be used full blood count (CBC) test example as the bone X ray of low hematocrit (anemia), low red blood cell count(RBC), low platelet and/or low numeration of leukocyte, bone marrow biopsy, serum protein electrophoresis, evaluation fracture and gets out osseous lesion thing or detect chemistry spectrum or the renal dysfunction that serum calcium, gross protein increase.

For example, can pass through the process of monitored in various ways renal cell carcinoma by doctor or veterinary, and can change thus dosage.The method of monitoring renal cell carcinoma comprises abdominal touch, full blood count (CBC), the experiment that detects erythrocytic urine examination, the rising of detection serum calcium level that for example detects bleb piece, experiment, urine cytology experiment, liver function test, abdomen and renal ultrasonography, kidney X ray, vein pyelograph (IVP) or the renal arteriography sheet that detects serum glutamic pyruvic transminase (SGPT) and alkali phosphatase rising.

The compositions and methods of the invention comprise IGF1R inhibitor optionally with one or more chemotherapeutics " coupling or combine ".Term " coupling or associating " refers to that the component of combination product of the present invention can be mixed with the single compositions for send simultaneously, or is mixed with respectively two or more compositionss (for example medicine box).In addition the time that, gives each component of curee's combination product of the present invention can be different from the time that gives other component; For example, each administration can be carried out (for example separately or sequential) within preset time simultaneously by some intervals are non-.And, can for example, by identical or different approach (oral, intravenous, subcutaneous), give curee independent component.

Embodiment

The present invention illustrates the present invention with following embodiment, rather than restriction the present invention.Below disclosed any method or compositions all fall within the scope of the present invention.

embodiment 1: in incidence cancerous cell, express IGF1R and IGF-II.

In the present embodiment, incidence cancerous cell has been carried out analyzing to measure the expression of IGF1R or IGF-2.Analyzed the IGF1RRNA expression of primary tumor tissue sample (deriving from 2 phases, 3 phases or 4 phase cancers), near normal tissue sample and normal structure sample.In addition also analyzed, the level of IGF1R, IGF-1 and IGF-2 in various cell line.

By tumor sample, prepare RNA, and synthesized cDNA by it.Adopt ABI Prism 7700 sequence detection system 9 (Applied Biosystems; Foster City, CA), in fluorogen 5 '-nuclease PCR experiment, use specific probe and primer, with 20ng cDNA sample, IGF1R is expressed and analyzed.By the CT number of all samples for ubiquitin or the normalization of actin mrna expression.

The primer and probe are as follows:

IGF1R/ forward primer: GAAAGTGACGTCCTGCATTTCA (SEQ ID NO:106)

IGF1R/ reverse primer: CCGGTGCCAGGTTATGATG (SEQ ID NO:107)

Probe sequence: CACCACCACGTCGAAGAATCGCA (SEQ ID NO:108)

H322 is Lines; CAL27, SCC25, SCC15, SCC9 and HS802 are squamous cell cancerous cell lines.

The data that obtain in these methods are shown in Fig. 1 and Fig. 2.Fig. 1 confirms in constitutional tumor of head and neck sample with the horizontal expression IGF1R higher than normal structure.And near the IGF1R expression of normal tissue (NAT) is greater than the level of normal structure.During the upgrading of tumor of head and neck stadium (by I to IV), the expression of IGF1R level is slightly improved.Each point has represented the normalization level of IGF1R in single tissue sample.

Fig. 2 has confirmed that IGF1R, IGF-I and IGF-II express in various lungs and squamous cell cancerous cell line equally.Upper figure is the western blot analysis of several cell lines, shows the expression of total IGF1R (tIGF-1R) in cell.Expression of actin also can be used as internal contrast.Figure below represents from the IGF1 of tested emiocytosis and IGF2 level (protein/106 cell of ng secretion).

embodiment 2: the propagation of anti-IGF1R (LCF (κ)/HCA (γ 1)) vitro inhibition squamous cell cancerous cell line

The present embodiment has confirmed that anti-IGF1R antibody LCF/HCA suppresses the propagation of different squamous cell carcinomas (SCC) cell line.

Application Cell-Titer Glo experiment (Promega Corp.; Madison, WI) measured cell proliferation.Under ATP from living cells exists, Cell-titer glo experiment produces " aura " signal that fluoresces, and this can use reads plate instrument luminometer (plate reader luminometer) or CCD imaging device detects.

In this experiment, squamous cell cancerous cell SCC9, SCC25 and SCC15 have been carried out to trypsinization, counting, and with 25,000 cell/ml Eddy diffusions in the 10%HI-FBS RPMI that contains NEAA, L-Glu, MEM vitamin and PS.100ml cell suspension (2500 cells) is added to BD Falcon in each hole of clear bottom black 96 orifice plates of TC processing.Make cell attachment, and breed and spend the night at 37 ℃.Next day, 10%RPMI culture medium is replaced with the 100ml 2%RPMI of the LCF/HCA antibody that contains suitable concn.Anti-IGF1R antibody concentration used in every experiment is 100nM, 20nM, 5nM, 0.8nM, 0.16nM, 0.032nM, 0.0064nM, 0.00128nM and 0.000256nM.In 2%RPMI, with 20 times of concentration, prepare all treatment samples, and carry out serial dilution.Each testing site is by preparing in independent assay plate in triplicate.As mentioned above, in treatment latter 96 hours, application CellTiter-Glo luminescent cell vitality test method (CellTiter-Glo Luminescent CellViability Assay) (Promega) on cell proliferation was measured.With being furnished with, be subject to the Wallac 420 of plate box (stacker) to read plate instrument to detect luminous.

This research the results are shown in Figure 3a-3c, in figure, show that the propagation of squamous cell cancerous cell line is suppressed (in figure, with 19D12, representing) by being exposed to the LCF/HCA of variable concentrations.Figure (a), figure (b) and to scheme above-mentioned cell line corresponding to the data of (c) as follows: (a): cell line SCC 15; (b): cell line SCC 25; (c): cell line SCC 9.

embodiment 3: IGF2 and IGF1R be high expressed in gastric cancer and ovarian cancer cell line.

In the present embodiment, confirmed to compare with normal cell system with normal specimens, IGF2mRNA expresses with higher level in Primary Gastric tumor sample stomach function regulating gland cell system.In tumor sample, the high expressed of IGF2 is by the labelling of the autocrine stimulation of IGF1R growth pathway, represents can suppress tumor growth by anti-IGF1R (LCF (κ)/HCA (γ 1)) Antybody therapy.

Substantially growth, preparation and the mensuration of the cell line of expression to be determined by method as mentioned above, have been carried out.By Western blotting, protein level is analyzed, by Taqman analytic process, mRNA level is analyzed.Application Bangs Laboratories, Inc. quantitative simple cell system (Quantitative Simply Cellular system), use the cell with anti-IGF1R antibody, anti-IR (Insulin receptor INSR) antibody, anti-EGFR (EGF-R ELISA) antibody or Anti-HER 2 labelling, carried out quantitative flow cytometry (Fishers, IN).

A2780 is Proliferation of Human Ovarian Cell system, and NCI-N87, SNU-16, SNU-1 and Hs746T are SGC-7901.

This research the results are shown in Figure 4 and Fig. 5.Fig. 4 has confirmed to compare with near normal cell with normal cell, and IGF2 expresses with higher level in Primary Gastric tumor cell.Each point is the normalization level (for the expression normalization of ubiquitin) at the IGF2mRNA of single neoplasmic tissue sample expression.

Upper figure in Fig. 5 is the quantitative flow cytometry data of relevant several cell lines, represents the receptor/cell number for IGF1R, IR, EGFR and HER2.Figure below in Fig. 5 is the Western blotting data of several cell lines, represents the expression of pIRS-1 (phosphorylation IRS-1 (IRS-1)), pAKT (phosphorylation AKT (also claiming protein kinase-B) (non-isotype specificity)), tIRS-1 (total IRS-1), pERK1, pERK2 (phosphorylation ERK1 (extracellular signal-regulated kinase 1) and ERK2 (extracellular signal-regulated kinase 2)), tIGR1R (total IGFR1) and actin.The migration of the band in t IRS-1 row NCI N87 swimming lane 3 is slower than the summary of expection, and this has shown that it is exactly the HER2 with anti-IRS-1 antibody generation cross reaction for this experiment.

embodiment 4: anti-IGF1R (LCF (κ)/HCA (γ 1)) antibody in vitro suppresses the propagation of gastric cancer and myeloma cell line.

The present embodiment has confirmed that anti-IGF1R antibody LCF/HCA suppresses the propagation of gastric carcinoma cell lines SNU-16 cell and myeloma cell line RPMI8226.Substantially press method as mentioned above, Growth of Cells, preparation and mensuration have been carried out in application Cell-Titer Glo experiment.Anti-IGF1R antibody concentration for every experiment is 100nm, 20nm, 5nm, 0.8nm, 0.16nm, 0.032nm, 0.0064nm, 0.00128nm and 0.000256nM.

This research the results are shown in Figure 6.Fig. 6 shows that the growth in vitro of SNU-16 cell line under the antibody of several concentration suppresses.The growth in vitro of also observing myeloma cell line 40%-45% level suppresses.

embodiment 5: anti-IGF1R (LCF (κ)/HCA (γ 1)) antibody suppresses renal cell carcinoma in vivo, suppresses in vitro the growth of melanoma tumor cell.

The present embodiment has confirmed to be used for the treatment of or to prevent the effect of renal cell carcinoma or melanomatous anti-IGF1R therapy.

By A498 people's renal carcinoma tumor cell and matrigel (Matrigel) (1: 1 cell: be inoculated into athymic nude mice right flank gel) subcutaneous.In these experiments, by 5x10 ⁶individual cell/mice is by with conventional matrigel mixing at 1: 1.The anti-IGF1R antibody of LCF/HCA is inoculated into intraperitoneal (ip) (0.1mg/ injection).By peritoneal injection, give mouse antibodies twice weekly, since the 12nd day totally 7 times.With caliper, measure tumor size, data typing LABCAT experimental data is gathered and analysis programme (Innovative Programming Associates, Inc.; Princeton, NJ).By mean size, be 100mm ³mice group.Measure tumor size and body weight twice weekly.

External test has been carried out in the reaction that is also the anti-IGF1R antibody of 100nM, 20nM, 5nM, 0.8nM, 0.16nM, 0.032nM, 0.0064nM, 0.00128nM and 0.000256nM to human melanoma cell (A375-SM) to concentration.By Cell-Titer Glo algoscopy, by method as mentioned above, measure.

This research the results are shown in Figure 7 and Fig. 8.Fig. 7 represents that temporal evolution suppresses the tumor growth of renal cell carcinoma cell line when giving LCF/HCA antibody.Fig. 8 is illustrated under different LCF/HCA antibody concentration the growth in vitro of melanoma cell series is suppressed.

embodiment 6: in anti-IGF1R (LCF (κ)/HCA (γ 1)) antibody body, suppress squamous cell growth of cancer cells.

The present embodiment has confirmed to be used for the treatment of or to prevent the effect of the anti-IGF1R therapy of incidence cancer (for example squamous cell carcinoma).

By SCC15 people's squamous cell carcinoma tumor cell and matrigel (1: 1 cell: be inoculated into immunodeficiency type SCID mice (strain SCID) right flank gel) subcutaneous.In these experiments, by 5x10 ⁶individual cell/mice mixed rear inoculation by 1: 1 with conventional matrigel.The anti-IGF1R antibody of LCF/HCA is inoculated into intraperitoneal (ip) (0.1mg/ injection).By peritoneal injection, give mouse antibodies twice weekly, since the 10th day totally 7 times, last administration in the time of the 31st day.With caliper, measure tumor size, by data typing labcat program.By mean size, be 100mm ³mice group.Measure tumor size and body weight twice weekly.

The data that obtained by these experiments are shown in Fig. 9.Inoculated the tumor growth (representing with " 19D12 " in figure) of the mice of LCF/HCA antibody than only accepting without viewed slow in antibody control injection mice.

embodiment 7: the PPTP group test of the anti-IGF1R antibody of LCF (κ)/HCA (γ 1).

LCF/HCA is the fully human antibodies for type-1 insulin like growth factor receptor (IGF1R), and it means growth and the transitivity phenotype of malignant tumor on a large scale.IGF1R signal transduction is particular importance under child's cancerous condition, for example with department of pediatrics cancer for example neuroblastoma, Ewing sarcoma, rhabdomyosarcoma, wilms' tumor (Wilms tumor) is relevant with osteosarcoma.In vitro and in vivo group antagonist activity for test program before Pediatric Clinic (Pediatric Preclinical Testing Program, PPTP) is evaluated.PPTP comprises by National Cancer Institute (National Cancer Institute, NCI) create body outer cell line group and the interior mice xenograft of the body group setting up, for the therapy of the modal department of pediatrics cancer of test (referring to such as Houghton etc., Pediatric Blood & Cancer (2007) 49 (7): 928-40).

PPTP comprises the He Tinei of body outer cell line group (n=27) the xenograft group (n=61) characterizing from molecule, and they have represented the modal type of Children with Solid Tumors and child ALL (acute lymphoblast leukemia).In the concentration range of 0.01nM～100nM, for the external group of PPTP, the anti-IGF1R antibody of LCF/HCA is measured; And at dosage, be 0.5mg/ mice, by peritoneal injection, twice administration reaches surrounding weekly in the situation that, for PPTP Ti Nei group, measure.Anti-tumor activity has been carried out to three times to be measured: 1) after clinical orientation (clinical setting), set reaction normal; 2) time treatment in the 21st day and the gross tumor volume that contrasts (T/C); With 3) according to the median EFS minute/event for the treatment of system and control series (gross tumor volume increases by 4 times) (medium activity needs EFS (Event-free survival rate (Event-freesurvival)) T/C > 2, high activity in addition need to be when experiment end median gross tumor volume only reduce).

Method

external test.Application DIMSCAN carries out external test, DIMSCAN is a kind of semi-automatic digital image microscope system based on fluorescence (using diacetic acid fluorescein [FDA] in tissue culture's porous plate) (Keshelava etc., Methods Mol.Med. (2005) 110:139-153) of quantitative assay viable count.Under the concentration from 0.01nM to 0.1mM, carried out the mensuration of 96 hours, 6 of every data points are duplicate.Application Kaleidagraph software (Synergy Software; Reading, PA), nonlinear regression and fitting, for reaction S shape dose-response model, relative fluorescence value and concentration, data are analyzed.The external group of PPTP comprises neuroblastoma (4), Ewing sarcoma (4), rhabdomyosarcoma (4), acute lymphoblast leukemia (5), NHL (2) cell line etc.

1 phase was measured to relate to and uses adult's preclinical models, according to PK/PD (pharmacokinetics/pharmacodynamics) research, be determined at its maximum tolerated dose (MTD) or under selected dosage, spread all over the medicine of the whole PPTP group of child's cancer xenograft system.

in body, entity tumor is measured: for each xenotransplantation system, when gross tumor volume is between 0.2-0.5cm ³between time, start treating with 10 mices of subcutaneous (SC) tumor.With digital caliper, press weekly two vertical diameter of tumor of interval measurement.Suppose that tumor is spherical, by formula (π/6) * d3, calculate volume, wherein d represents average diameter.

in body, acute lymphoblast leukemia is measured: for each xenotransplantation system, with the 3-5x10 of being purified out by the spleen of Secondary cases receptor (secondary recipient) mice ⁶individual mononuclear cell is given 8 mouse inoculations.With flow cytometry, monitor weekly implantation situation, people CD45 in peripheral blood ⁺the begin treatment when ratio of cell reaches 1%.People CD45 in monitoring peripheral blood weekly in whole therapeutic process ⁺the ratio of cell.

medicine: LCF/HCA is dissolved in to 20mM sodium acetate (pH5) buffer that contains 150mM sodium chloride, by the dosage of 0.5mg/ animal through intraperitoneal twice continuous surrounding administration weekly.According to PPTP S.O.P., the antibody in numbering bottle is supplied with to each and measure position for Blind Test.

entity tumor reaction normal:

Reaction	Definition	Scoring
			PD1 (PD 1)	Gross tumor volume ↑ > 25%, TGD value≤1.5	0
PD2 (PD 2)	Gross tumor volume ↑ > 25%, TGD value > 1.5	2
			SD (stability disease)	Gross tumor volume ↑ < 25%, and < 50% disappears	4
PR (partial reaction)	Disappear >=50%, but non-CR	6
			CR (complete reaction)	Gross tumor volume < 0.1cm 3	8
MCR (maintaining CR)	Gross tumor volume < 0.1cm when research finishes 3	10

leukemia reaction normal:

Reaction	Definition	Scoring
			PD1 (PD 1)	Without PR, TGD value≤1.5, the event of EOS aspect	0
PD2 (PD 2)	Without PR, TGD value > 1.5, the event of EOS aspect	2
			SD (stability disease)	Without PR, without the event of EOS aspect	4
PR (partial reaction)	CD45% < 1% only one week	6
			CR (complete reaction)	Continuous 2 weeks of CD45% < 1%	8
MCR (maintaining CR)	CD45% < 1% until research last 3 weeks	10

the reaction of median group: give every mice reaction scoring (ginseng sees the above table) in each treatment group, calculate the median scoring for the treatment of group, then according to following table, determine the general reaction of each treatment group.

If the scoring of the median (1) (MS):	Overall Group's reaction
		0≤MS≤1	PD1
1＜MS≤3	PD2
		3＜MS≤5	SD
5＜MS≤7	PR
		7＜MS≤9	CR
9＜MS	MCR

statistical method: apply accurate log rank check, the Event-free survival rate (EFS) of each treatment group is distributed and contrasted with the EFS distribution of corresponding matched group.P-value is both sides, for this, studies given exploration character, multiple comparisons is not adjusted.P-value < 0.05 is regarded as significantly.

Result

Table 1.LCF/HCA activity in vivo gathers

* p value hurdle represents there is statistical significance between treatment group and matched group.The symbol on EFS hurdle represent to there is high activity (! ), the xenograft of medium activity (#) or uncertain activity (^).RTV is relative tumour volume.

Separately, referring to Figure 10, represent the time dependent graphic analysis of in-vivo tumour volume of different cell types.

Separately with Affymetrix U133Plus2 array, IGF-1R is expressed and evaluated.

IGF1R expresses not antagonist react (being PD1) of low-down 5 xenografts.For example, the expression of OS-33 is very low, is in aitiogenic two the osteosarcoma xenografts of antagonist not.The expression of 3 xenograft IGF1R with CR or MCR reaction is very high.In having, wait until not antagonist (for example Rh41) the generation reaction of some xenografts that high IGF1R expresses.

It is 30% that the maximum growth reaching in external test method suppresses, and has observed T cell ALL system, MOLT-4.Under the highest test concentrations, the median growth inhibited of external group is 5%.

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Scope of the present invention is not subject to the restriction of specific embodiments described herein.In fact, scope of the present invention comprises specific embodiments disclosed and herein concrete disclosed other embodiment herein; Specific embodiments disclosed exhaustive not necessarily herein.From aforementioned description, except content described herein, various modifications of the present invention are all apparent to those skilled in the art.This class is revised and is also all fallen in the scope of appended claims.

In the application's full text, quoted patent, patent application, publication, product description and scheme, its disclosure is all attached to by reference herein for all objects.

Claims (12)

1. Separation antibody for the preparation for the treatment of or prevention mammalian subject the medicine that is selected from following medical conditions in purposes, this antibody comprises light chain immune globulin variable region and heavy chain immunoglobulin variable region, the aminoacid 20-128 that described light chain immune globulin variable region comprises SEQ ID NO:8, is connected with κ constant region for immunoglobulin; The aminoacid 20-137 that described heavy chain immunoglobulin variable region comprises SEQ ID NO:10, be connected with γ-1 constant region for immunoglobulin, described medical conditions is selected from: incidence cancer, squamous cell carcinoma, gastric cancer, ovarian cancer, myeloma, renal cell carcinoma and melanoma.
2. 2. the purposes of claim 1, wherein said medical conditions is incidence cancer.
3. 3. the purposes of claim 1, the chemotherapeutics that wherein said antibody is other with one or more or its pharmaceutical composition are combined and are given.
4. 4. the purposes of claim 3, wherein said other chemotherapeutics is anticarcinogen, Bendectin, anti-anemic drug or anti-mucositis medicine.
5. 5. the purposes of claim 3, wherein said other chemotherapeutics is that one or more are selected from following medicine: everolimus; ET-743; Effect of nano-paclitaxel injection suspension; Telcyta; Ficlatuzumab; Pazopanib; GSK690693; RTA744; ON0910.Na; AZD6244 (ARRY-142886); AMN-107; TKI-258; GSK461364; AZD1152; Grace is pricked cry loudly woods; ZD6474; ARQ-197; MK-0457; MLN8054; PHA-739358; R-763; AT-9263; FLT-3 inhibitor; VEGFR inhibitor; EGFRTK inhibitor; Aurora inhibitors of kinases; PIK-1 regulator; Bcl-2 inhibitor; Hdac inhibitor; Inhibitors of c-met; PARP inhibitor; Cdk inhibitor; EGFR TK inhibitor; IGFR-TK inhibitor; Anti-HGF antibody; PI3 inhibitors of kinases; AKT inhibitor; JAK/STAT inhibitor; Restriction point-1 inhibitor; Restriction point-inhibitor 2; Focal adhesion kinase inhibitor; Map kinase kinase (mek) inhibitor; VEGF trap antibody; Pemetrexed; Erlotinib; Dasatinib; AMN107; Decitabine; Victibix; Amrubicin; Ao Gefu monoclonal antibody; Lep-etu; 2-Amino-6-methyl-5-(pyridin-4-ylsulfanyl)-3H-quinazolin-4-one; Azd2171; Ba Tabulin; Method wood monoclonal antibody difficult to understand; Prick wooden monoclonal antibody; Ai Te click woods; Tetrandrine; Rubitecan; Tesmilifene; Ao Limeisheng; For western wooden monoclonal antibody; Her wooden monoclonal antibody; Gossypol; Bio111; 131-I-TM-601; ALT-110; BIO140; CC8490; Cilengitide; Gefitinib; IL13-PE38QQR; INO1001; IPdR; KRX-0402; Lucanthone; LY317615; Iodine [ ¹³¹i] anti-tenascin monoclonal antibody 81C6; Vitespan; Rta744; Sdx102; Talampanel; Atrasentan; Xr311; Sieve meter is new; ADS-100380;

   

   cG-781; CG-1521;

   

   sB-556629; Chlamydocin; JNJ-16241199;

   

   

   

   SAHA; Etoposide; Gemcitabine; Doxorubicin; Liposome doxorubicin; 5 '-'-Deoxy-5-fluorouridine; Vincristine; Temozolomide; ZK-304709; Fill in sharp Seeley; PD0325901; AZD-6244; Capecitabine; Pidolidone; N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo-[2,3-d] pyrimidine-5-yl) ethyl] benzoyl]-disodium salt heptahydrate; Camptothecine; Irinotecan; Irinotecan, 5-fluorouracil and folinic acid combination product; The irinotecan of PEG labelling; FOLFOX scheme; Tamoxifen; Toremifene Citrate; Anastrozole; Exemestane; Letrozole; DES (diethylstilbestrol); Estradiol; Estrogen; Conjugated estrogen hormone; Bevacizumab; IMC-1C11; CHIR-258;

   

   3-[5-(methyl sulphonyl piperidine methyl)-indyl]-quinolinones; Vatalanib; AG-013736; AVE-0005; [D-Ser (But) 6, Azgly10]

   (pyro-Glu-His-Trp-Ser-Tyr-D-Ser (But)-Leu-Arg-Pro-Azgly-NH ₂acetate [C ₅₉h ₈₄n ₁₈o ₁₄(C ₂h ₄o ₂) _x; X=1-2.4 wherein]; Goserelin acetate; Acetic acid leuproside; Triptorelin pamoate; Sutent; Sunitinib malate; Medroxyprogesterone acetate; Hydroxyprogesterone caproate; Megestrol acetate; Raloxifene; Bicalutamide; Flutamide; Nilutamide; Megestrol acetate; CP-724714; TAK-165; HKI-272; Erlotinib; Lapatinib; Ka Na is for Buddhist nun; ABX-EGF antibody; Erbitux; EKB-569; PKI-166; GW-572016; Chlorine Na Fani;

   

   bMS-214662; Zarnestra; Amifostine; NVP-LAQ824; Suberoylanilide hyroxamic acid; Valproic acid; Trichostatin A; FK-228; SU11248; Sorafenib; KRN951; Aminoglutethimide; Amsacrine; Anagrelide; L-ASP; Bacillus calmette-guerin vaccine (BCG) vaccine; Bleomycin; Buserelin; Busulfan; Carboplatin; Carmustine; Chlorambucil; Cisplatin; Cladribine; Clodronate; Cyclophosphamide; Cyproterone; Cytosine arabinoside; Dacarbazine; Actinomycin D; Daunorubicin; Diethylstilbestrol; Epirubicin; Fludarabine; Fludrocortisone; Fluoxymesterone; Flutamide; Hydroxyurea; Idarubicin; Ifosfamide; Imatinib; Folinic acid; Leuproside; Levamisole; Lomustine; Chlormethine; Melphalan; Ismipur; Mesna; Methotrexate; Mitomycin; Mitotane; Mitoxantrone; Nilutamide; Octreotide; Ao Shalipa; Satraplatin pamldronate; Pentostatin; Plicamycin; Porphin nurse; Procarbazine; Raltitrexed; Rituximab; Streptozocin; Teniposide; Testosterone; Thalidomide; Thioguanine; Phosphinothioylidynetrisaziridine; Tretinoin; Vindesine; 13-cisRA; Melphalan; Uracil mustard; Estramustine; Altretamine; Floxuridine; 5-FU; Cytosine arabinoside; Ismipur; Deoxycoformycin; Calcitriol; Valrubicin; Mithramycin; Vinblastine; Vinorelbine; Hycamtin; Razoxin; Marimastat; COL-3; Neovastat; BMS-275291; Amine is overstated by department; Endostatin; SU5416; SU6668; EMD121974; IL-12; IM862; Angiostatin; The anti-monoclonal antibody of α V β 3 humanization; Droloxifene; Indoxifene; Spironolactone; Finasteride; Cimetidine; Herceptin; Denileukin diphtheria toxin, diphtherotoxin transmembrane segment; Gefitinib; Bortezomib; Paclitaxel; Agalactia floats paclitaxel; Docetaxel; Epothilone B; BMS-247550; BMS-310705; Droloxifene; 4-hydroxytamoxifen; ERA 923; ERA-923; Arzoxifene; Fulvestrant; Acolbifene; Lasofoxifene; Idoxifene; TSE-424; HMR-3339; ZK186619; Hycamtin; PTK787/ZK222584; VX-745; PD184352; Rapamycin; 40-O-(2-hydroxyethyl)-rapamycin; CCI-779; AP-23573; RAD001; ABT-578; BC-210; LY294002; LY292223; LY292696; LY293684; LY293646; Wortmannin; ZM336372; L-779,450; PEG-filgrastim; Darbepoetin; 5-fluorouracil; Erythropoietin; Granulocyte colony-stimulating factor; Zoledronate; Prednisone; Cetuximab; Granulocyte macrophage colony stimulating factor; Histrelin; Glycol interferon alpha-2a; Intederon Alpha-2a; Glycol interferon alpha-2b; Interferon Alpha-2b; Azacitidine; PEG-L-asparaginase; Lenalidomide; Lucky trastuzumab; Hydrocortisone; Interleukin-11; Dexrazoxane; A Lun pearl monoclonal antibody; All-trans-retinoic acid; Ketoconazole; Interleukin-2; Megestrol; Immunoglobulin; Chlormethine; Methylprednisolone; Ibritumomab tiuxetan; Androgen; Decitabine; Altretamine; Bexarotene; Tositumomab; Arsenic trioxide; Cortisone; Editronate; Mitotane; Cyclosporin; Liposome daunorubicin; Edwina-asparaginase; Strontium 89; Carcel is smooth; Netupitant; Nk 1 receptor antagonist; Palonosetron; Aprepitant; Diphenhydramine; Hydroxyzine; Metoclopramide; Lorazepam; Alprazolam; Haloperidol; Droperidol; Dronabinol; Dexamethasone; Methylprednisolone; Prochlorperazine; Granisetron; Ondansetron; Dolasetron; Tropisetron; Filgrastim; Erythropoietin; Epoetin Alfa; With darbepoetin α.
6. 6. the purposes of claim 3, wherein said antibody and other chemotherapeutics give simultaneously.
7. 7. the purposes of claim 3, wherein said antibody and other chemotherapeutics are non-to be given simultaneously.
8. 8. the purposes of claim 1, wherein said curee is people.
9. 9. the purposes of claim 8, wherein said curee is child.
10. 10. the purposes of claim 1, wherein said antibody is combined and is given with anti-cancer therapies.
11. The purposes of 11. claim 10, wherein said anti-cancer therapies is surgery tumorectomy and/or anticancer radiotherapy.
12. The purposes of 12. claim 1, wherein said antibody gives described curee with pharmaceutical compositions, described pharmaceutical composition comprises 1,5,10,15,20 or 25mg/ml antibody, 1.8,2.3 or 3.1mg/ml sodium acetate trihydrate, 0.5,0.18 or 2.2mg/ml glacial acetic acid, 50 or 70mg/ml sucrose and water, and pH is 5.5.

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